From c8fdd9b3776648d0c459cd0b0c0fc8bd5083c485 Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 1 Nov 2021 17:20:41 +0100 Subject: [PATCH 001/246] add aDNA workflow --- modules.json | 23 +++++- .../modules/bcftools/consensus/functions.nf | 78 ++++++++++++++++++ .../modules/bcftools/consensus/main.nf | 40 ++++++++++ .../modules/bcftools/consensus/meta.yml | 49 ++++++++++++ .../modules/bcftools/index/functions.nf | 78 ++++++++++++++++++ .../nf-core/modules/bcftools/index/main.nf | 44 ++++++++++ .../nf-core/modules/bcftools/index/meta.yml | 49 ++++++++++++ .../modules/bcftools/view/functions.nf | 78 ++++++++++++++++++ modules/nf-core/modules/bcftools/view/main.nf | 53 ++++++++++++ .../nf-core/modules/bcftools/view/meta.yml | 63 +++++++++++++++ .../modules/freebayes/germline/functions.nf | 78 ++++++++++++++++++ .../modules/freebayes/germline/main.nf | 80 +++++++++++++++++++ .../modules/freebayes/germline/meta.yml | 78 ++++++++++++++++++ .../modules/pydamage/analyze/functions.nf | 78 ++++++++++++++++++ .../nf-core/modules/pydamage/analyze/main.nf | 42 ++++++++++ .../nf-core/modules/pydamage/analyze/meta.yml | 55 +++++++++++++ .../modules/pydamage/filter/functions.nf | 78 ++++++++++++++++++ .../nf-core/modules/pydamage/filter/main.nf | 42 ++++++++++ .../nf-core/modules/pydamage/filter/meta.yml | 51 ++++++++++++ .../modules/samtools/faidx/functions.nf | 78 ++++++++++++++++++ .../nf-core/modules/samtools/faidx/main.nf | 36 +++++++++ .../nf-core/modules/samtools/faidx/meta.yml | 33 ++++++++ subworkflows/local/ancient_dna.nf | 35 ++++++++ 23 files changed, 1318 insertions(+), 1 deletion(-) create mode 100644 modules/nf-core/modules/bcftools/consensus/functions.nf create mode 100644 modules/nf-core/modules/bcftools/consensus/main.nf create mode 100644 modules/nf-core/modules/bcftools/consensus/meta.yml create mode 100644 modules/nf-core/modules/bcftools/index/functions.nf create mode 100644 modules/nf-core/modules/bcftools/index/main.nf create mode 100644 modules/nf-core/modules/bcftools/index/meta.yml create mode 100644 modules/nf-core/modules/bcftools/view/functions.nf create mode 100644 modules/nf-core/modules/bcftools/view/main.nf create mode 100644 modules/nf-core/modules/bcftools/view/meta.yml create mode 100644 modules/nf-core/modules/freebayes/germline/functions.nf create mode 100644 modules/nf-core/modules/freebayes/germline/main.nf create mode 100644 modules/nf-core/modules/freebayes/germline/meta.yml create mode 100644 modules/nf-core/modules/pydamage/analyze/functions.nf create mode 100644 modules/nf-core/modules/pydamage/analyze/main.nf create mode 100644 modules/nf-core/modules/pydamage/analyze/meta.yml create mode 100644 modules/nf-core/modules/pydamage/filter/functions.nf create mode 100644 modules/nf-core/modules/pydamage/filter/main.nf create mode 100644 modules/nf-core/modules/pydamage/filter/meta.yml create mode 100644 modules/nf-core/modules/samtools/faidx/functions.nf create mode 100644 modules/nf-core/modules/samtools/faidx/main.nf create mode 100644 modules/nf-core/modules/samtools/faidx/meta.yml create mode 100644 subworkflows/local/ancient_dna.nf diff --git a/modules.json b/modules.json index 5caaeb81..23933cfe 100644 --- a/modules.json +++ b/modules.json @@ -3,15 +3,36 @@ "homePage": "https://github.com/nf-core/mag", "repos": { "nf-core/modules": { + "bcftools/consensus": { + "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + }, + "bcftools/index": { + "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + }, + "bcftools/view": { + "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + }, "fastp": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, "fastqc": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, + "freebayes/germline": { + "git_sha": "3df4fe60851c19ec8d55ef3a978ae851eaccfd68" + }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" + }, + "pydamage/analyze": { + "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" + }, + "pydamage/filter": { + "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" + }, + "samtools/faidx": { + "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" } } } -} +} \ No newline at end of file diff --git a/modules/nf-core/modules/bcftools/consensus/functions.nf b/modules/nf-core/modules/bcftools/consensus/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/bcftools/consensus/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/bcftools/consensus/main.nf b/modules/nf-core/modules/bcftools/consensus/main.nf new file mode 100644 index 00000000..954b0eb8 --- /dev/null +++ b/modules/nf-core/modules/bcftools/consensus/main.nf @@ -0,0 +1,40 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process BCFTOOLS_CONSENSUS { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' + } else { + container 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' + } + + input: + tuple val(meta), path(vcf), path(tbi), path(fasta) + + output: + tuple val(meta), path('*.fa'), emit: fasta + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + """ + cat $fasta | bcftools consensus $vcf $options.args > ${prefix}.fa + header=\$(head -n 1 ${prefix}.fa | sed 's/>//g') + sed -i 's/\${header}/${meta.id}/g' ${prefix}.fa + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/bcftools/consensus/meta.yml b/modules/nf-core/modules/bcftools/consensus/meta.yml new file mode 100644 index 00000000..761115a6 --- /dev/null +++ b/modules/nf-core/modules/bcftools/consensus/meta.yml @@ -0,0 +1,49 @@ +name: bcftools_consensus +description: Compresses VCF files +keywords: + - variant calling + - consensus + - VCF +tools: + - consensus: + description: | + Create consensus sequence by applying VCF variants to a reference fasta file. + homepage: http://samtools.github.io/bcftools/bcftools.html + documentation: http://www.htslib.org/doc/bcftools.html + doi: 10.1093/bioinformatics/btp352 + licence: ['MIT'] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: VCF file + pattern: "*.{vcf}" + - tbi: + type: file + description: tabix index file + pattern: "*.{tbi}" + - fasta: + type: file + description: FASTA reference file + pattern: "*.{fasta,fa}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: FASTA reference consensus file + pattern: "*.{fasta,fa}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" diff --git a/modules/nf-core/modules/bcftools/index/functions.nf b/modules/nf-core/modules/bcftools/index/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/bcftools/index/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/bcftools/index/main.nf b/modules/nf-core/modules/bcftools/index/main.nf new file mode 100644 index 00000000..d67614d8 --- /dev/null +++ b/modules/nf-core/modules/bcftools/index/main.nf @@ -0,0 +1,44 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process BCFTOOLS_INDEX { + tag "$meta.id" + label 'process_low' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0" + } else { + container "quay.io/biocontainers/bcftools:1.13--h3a49de5_0" + } + + input: + tuple val(meta), path(vcf) + + output: + tuple val(meta), path("*.csi"), optional:true, emit: csi + tuple val(meta), path("*.tbi"), optional:true, emit: tbi + path "versions.yml" , emit: version + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + + """ + bcftools \\ + index \\ + $options.args \\ + --threads $task.cpus \\ + $vcf + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/bcftools/index/meta.yml b/modules/nf-core/modules/bcftools/index/meta.yml new file mode 100644 index 00000000..0d5dd3ef --- /dev/null +++ b/modules/nf-core/modules/bcftools/index/meta.yml @@ -0,0 +1,49 @@ +name: bcftools_index +description: Index VCF tools +keywords: + - vcf + - index + - bcftools + - csi + - tbi +tools: + - bcftools: + description: BCFtools is a set of utilities that manipulate variant calls in the Variant Call Format (VCF) and its binary counterpart BCF. All commands work transparently with both VCFs and BCFs, both uncompressed and BGZF-compressed. Most commands accept VCF, bgzipped VCF and BCF with filetype detected automatically even when streaming from a pipe. Indexed VCF and BCF will work in all situations. Un-indexed VCF and BCF and streams will work in most, but not all situations. + homepage: https://samtools.github.io/bcftools/ + documentation: https://samtools.github.io/bcftools/howtos/index.html + tool_dev_url: https://github.com/samtools/bcftools + doi: "10.1093/gigascience/giab008" + licence: ['MIT', 'GPL-3.0-or-later'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - VCF: + type: file + description: VCF file (optionally GZIPPED) + pattern: "*.{vcf,vcf.gz}" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - version: + type: file + description: File containing software version + pattern: "versions.yml" + - csi: + type: file + description: Default VCF file index file + pattern: "*.csi" + - tbi: + type: file + description: Alternative VCF file index file for larger files (activated with -t parameter) + pattern: "*.tbi" + +authors: + - "@jfy133" diff --git a/modules/nf-core/modules/bcftools/view/functions.nf b/modules/nf-core/modules/bcftools/view/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/bcftools/view/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/bcftools/view/main.nf b/modules/nf-core/modules/bcftools/view/main.nf new file mode 100644 index 00000000..ef72f081 --- /dev/null +++ b/modules/nf-core/modules/bcftools/view/main.nf @@ -0,0 +1,53 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process BCFTOOLS_VIEW { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::bcftools=1.13" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0" + } else { + container "quay.io/biocontainers/bcftools:1.13--h3a49de5_0" + } + + input: + tuple val(meta), path(vcf), path(index) + path(regions) + path(targets) + path(samples) + + output: + tuple val(meta), path("*.gz") , emit: vcf + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def regions_file = regions ? "--regions-file ${regions}" : "" + def targets_file = targets ? "--targets-file ${targets}" : "" + def samples_file = samples ? "--samples-file ${samples}" : "" + + + """ + bcftools view \\ + --output ${prefix}.vcf.gz \\ + ${regions_file} \\ + ${targets_file} \\ + ${samples_file} \\ + $options.args \\ + --threads $task.cpus \\ + ${vcf} + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/bcftools/view/meta.yml b/modules/nf-core/modules/bcftools/view/meta.yml new file mode 100644 index 00000000..df5b0f8f --- /dev/null +++ b/modules/nf-core/modules/bcftools/view/meta.yml @@ -0,0 +1,63 @@ +name: bcftools_view +description: View, subset and filter VCF or BCF files by position and filtering expression. Convert between VCF and BCF +keywords: + - variant calling + - view + - bcftools + - VCF + +tools: + - view: + description: | + View, subset and filter VCF or BCF files by position and filtering expression. Convert between VCF and BCF + homepage: http://samtools.github.io/bcftools/bcftools.html + documentation: http://www.htslib.org/doc/bcftools.html + doi: 10.1093/bioinformatics/btp352 + licence: ['MIT'] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: | + The vcf file to be inspected. + e.g. 'file.vcf' + - index: + type: file + description: | + The tab index for the VCF file to be inspected. + e.g. 'file.tbi' + - regions: + type: file + description: | + Optionally, restrict the operation to regions listed in this file. + e.g. 'file.vcf' + - targets: + type: file + description: | + Optionally, restrict the operation to regions listed in this file (doesn't rely upon index files) + e.g. 'file.vcf' + - samples: + type: file + description: | + Optional, file of sample names to be included or excluded. + e.g. 'file.tsv' +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: VCF normalized output file + pattern: "*.{vcf.gz}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@abhi18av" diff --git a/modules/nf-core/modules/freebayes/germline/functions.nf b/modules/nf-core/modules/freebayes/germline/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/freebayes/germline/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/freebayes/germline/main.nf b/modules/nf-core/modules/freebayes/germline/main.nf new file mode 100644 index 00000000..eae62036 --- /dev/null +++ b/modules/nf-core/modules/freebayes/germline/main.nf @@ -0,0 +1,80 @@ +// Import generic module functions +include { initOptions; saveFiles; getProcessName; getSoftwareName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process FREEBAYES_GERMLINE { + tag "$meta.id" + label 'process_low' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::freebayes=1.3.5" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/freebayes:1.3.5--py38ha193a2f_3" + } else { + container "quay.io/biocontainers/freebayes:1.3.5--py38ha193a2f_3" + } + + input: + tuple val(meta), path(input), path(input_index) + path fasta + path fai + path targets + path samples + path populations + path cnv + + output: + tuple val(meta), path("*.vcf.gz") , emit: vcf + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def targets_file = targets ? "--target ${targets}" : "" + def samples_file = samples ? "--samples ${samples}" : "" + def populations_file = populations ? "--populations ${populations}" : "" + def cnv_file = cnv ? "--cnv-map ${cnv}" : "" + + if (task.cpus > 1) { + """ + freebayes-parallel \\ + <(fasta_generate_regions.py ${fasta}.fai 10000) ${task.cpus} \\ + -f $fasta \\ + $targets_file \\ + $samples_file \\ + $populations_file \\ + $cnv_file \\ + $options.args \\ + $input > ${prefix}.vcf + + gzip --no-name ${prefix}.vcf + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) + END_VERSIONS + """ + + } else { + """ + freebayes \\ + -f $fasta \\ + $targets_file \\ + $samples_file \\ + $populations_file \\ + $cnv_file \\ + $options.args \\ + $input > ${prefix}.vcf + + gzip --no-name ${prefix}.vcf + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) + END_VERSIONS + """ + } +} diff --git a/modules/nf-core/modules/freebayes/germline/meta.yml b/modules/nf-core/modules/freebayes/germline/meta.yml new file mode 100644 index 00000000..86650715 --- /dev/null +++ b/modules/nf-core/modules/freebayes/germline/meta.yml @@ -0,0 +1,78 @@ +name: freebayes_germline +description: A haplotype-based variant detector +keywords: + - variant caller + - SNP + - genotyping + - variant calling + - bayesian +tools: + - freebayes: + description: Bayesian haplotype-based polymorphism discovery and genotyping + homepage: https://github.com/freebayes/freebayes + documentation: https://github.com/freebayes/freebayes + tool_dev_url: https://github.com/freebayes/freebayes + doi: "arXiv:1207.3907" + licence: ['MIT'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - input_index: + type: file + description: BAM/CRAM/SAM index file + pattern: "*.bam.bai" + - fasta: + type: file + description: reference fasta file + pattern: ".{fa,fa.gz,fasta,fasta.gz}" + - fai: + type: file + description: reference fasta file index + pattern: "*.fai" + - targets: + type: file + description: Optional - Limit analysis to targets listed in this BED-format FILE. + pattern: "*.bed" + - samples: + type: file + description: Optional - Limit analysis to samples listed (one per line) in the FILE. + pattern: "*.txt" + - populations: + type: file + description: Optional - Each line of FILE should list a sample and a population which it is part of. + pattern: "*.txt" + - cnv: + type: file + description: | + A copy number map BED file, which has + either a sample-level ploidy: + sample_name copy_number + or a region-specific format: + seq_name start end sample_name copy_number + pattern: "*.bed" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - version: + type: file + description: File containing software version + pattern: "*.{version.txt}" + - vcf: + type: file + description: Compressed VCF file + pattern: "*.vcf.gz" +authors: + - "@maxibor" + - "@FriederikeHanssen" diff --git a/modules/nf-core/modules/pydamage/analyze/functions.nf b/modules/nf-core/modules/pydamage/analyze/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/pydamage/analyze/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/pydamage/analyze/main.nf b/modules/nf-core/modules/pydamage/analyze/main.nf new file mode 100644 index 00000000..9cfb8a1a --- /dev/null +++ b/modules/nf-core/modules/pydamage/analyze/main.nf @@ -0,0 +1,42 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process PYDAMAGE_ANALYZE { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0" + } else { + container "quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0" + } + + input: + tuple val(meta), path(bam), path(bai) + + output: + tuple val(meta), path("pydamage_results/pydamage_results.csv"), emit: csv + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + """ + pydamage \\ + analyze \\ + $options.args \\ + -p $task.cpus \\ + $bam + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/pydamage/analyze/meta.yml b/modules/nf-core/modules/pydamage/analyze/meta.yml new file mode 100644 index 00000000..918fbce9 --- /dev/null +++ b/modules/nf-core/modules/pydamage/analyze/meta.yml @@ -0,0 +1,55 @@ +name: pydamage_analyze +description: Damage parameter estimation for ancient DNA +keywords: + - ancient DNA + - aDNA + - de novo assembly + - filtering + - damage + - deamination + - miscoding lesions + - C to T + - palaeogenomics + - archaeogenomics + - palaeogenetics + - archaeogenetics +tools: + - pydamage: + description: Damage parameter estimation for ancient DNA + homepage: https://github.com/maxibor/pydamage + documentation: https://pydamage.readthedocs.io/ + tool_dev_url: https://github.com/maxibor/pydamage + licence: ['GPL v3'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - bai: + type: file + description: BAM/CRAM/SAM index file + pattern: "*.{bai,crai,sai}" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - csv: + type: file + description: PyDamage results as csv files + pattern: "*.csv" + +authors: + - "@maxibor" diff --git a/modules/nf-core/modules/pydamage/filter/functions.nf b/modules/nf-core/modules/pydamage/filter/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/pydamage/filter/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/pydamage/filter/main.nf b/modules/nf-core/modules/pydamage/filter/main.nf new file mode 100644 index 00000000..6cd7ae7a --- /dev/null +++ b/modules/nf-core/modules/pydamage/filter/main.nf @@ -0,0 +1,42 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process PYDAMAGE_FILTER { + tag "$meta.id" + label 'process_low' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0" + } else { + container "quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0" + } + + input: + tuple val(meta), path(csv) + + output: + tuple val(meta), path("pydamage_results/pydamage_filtered_results.csv"), emit: csv + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + """ + + pydamage \\ + filter \\ + $options.args \\ + $csv + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/pydamage/filter/meta.yml b/modules/nf-core/modules/pydamage/filter/meta.yml new file mode 100644 index 00000000..706e38b0 --- /dev/null +++ b/modules/nf-core/modules/pydamage/filter/meta.yml @@ -0,0 +1,51 @@ +name: pydamage_filter +description: Damage parameter estimation for ancient DNA +keywords: + - ancient DNA + - aDNA + - de novo assembly + - filtering + - damage + - deamination + - miscoding lesions + - C to T + - palaeogenomics + - archaeogenomics + - palaeogenetics + - archaeogenetics +tools: + - pydamage: + description: Damage parameter estimation for ancient DNA + homepage: https://github.com/maxibor/pydamage + documentation: https://pydamage.readthedocs.io/ + tool_dev_url: https://github.com/maxibor/pydamage + licence: ['GPL v3'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - csv: + type: file + description: csv file from pydamage analyze + pattern: "*.csv" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - csv: + type: file + description: PyDamage filtered results as csv file + pattern: "*.csv" + +authors: + - "@maxibor" diff --git a/modules/nf-core/modules/samtools/faidx/functions.nf b/modules/nf-core/modules/samtools/faidx/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/samtools/faidx/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/samtools/faidx/main.nf b/modules/nf-core/modules/samtools/faidx/main.nf new file mode 100644 index 00000000..80cedeab --- /dev/null +++ b/modules/nf-core/modules/samtools/faidx/main.nf @@ -0,0 +1,36 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process SAMTOOLS_FAIDX { + tag "$fasta" + label 'process_low' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:[:], publish_by_meta:[]) } + + conda (params.enable_conda ? 'bioconda::samtools=1.13' : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/samtools:1.13--h8c37831_0" + } else { + container "quay.io/biocontainers/samtools:1.13--h8c37831_0" + } + + input: + path fasta + + output: + path "*.fai" , emit: fai + path "versions.yml", emit: versions + + script: + """ + samtools faidx $fasta + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/samtools/faidx/meta.yml b/modules/nf-core/modules/samtools/faidx/meta.yml new file mode 100644 index 00000000..16c0b334 --- /dev/null +++ b/modules/nf-core/modules/samtools/faidx/meta.yml @@ -0,0 +1,33 @@ +name: samtools_faidx +description: Index FASTA file +keywords: + - index + - fasta +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ['MIT'] +input: + - fasta: + type: file + description: FASTA file + pattern: "*.{fa,fasta}" +output: + - fai: + type: file + description: FASTA index file + pattern: "*.{fai}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@ewels" + - "@phue" diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf new file mode 100644 index 00000000..7fe14559 --- /dev/null +++ b/subworkflows/local/ancient_dna.nf @@ -0,0 +1,35 @@ +include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main.nf' addParams( options: [:] ) +include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main.nf' addParams( options: [:] ) +include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main.nf' addParams( options: [:] ) +include { FREEBAYES_GERMLINE as FREEBAYES } from '../../modules/nf-core/modules/freebayes/germline/main.nf' addParams( options: [args : "-p 1"] ) +include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main.nf' addParams( options: [:] ) +include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main.nf' addParams( options: [:] ) +include { SAMTOOLS_FAIDX as FAIDX} from from '../../modules/nf-core/modules/samtools/faidx/main.nf' addParams( options: [:] ) + +worflow ANCIENT_DNA_ASSEMLY_VALIDATION { + take: + contigs // assembled contigs [ val(meta), [contigs] ] + bam // reads mapped back on contigs [ val(meta), [bam] ] + bam_index // bam index [ val(meta), [index] ] + main: + PYDAMAGE_ANALYZE(contigs + .join(bam) + .join(bam_index)) + PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) + FAIDX(contigs.map { it -> [ it[1] ] }) + FREEBAYES (bam.join(bam_index), contigs.map { it -> [ it[1] ] }, FAIDX.out.FAI ) + BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) + BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi)) // Add filtering parameters + BCFTOOLS_INDEX_POST(BCFTOOLS_VIEW.out.vcf) + BCFTOOLS_CONSENSUS(BCFTOOLS_VIEW.out.vcf + .join(BCFTOOLS_INDEX_POST.out.tbi) + .join(contigs)) + emit: + contigs_recalled: BCFTOOLS_CONSENSUS.out.fasta + pydamage_results: PYDAMAGE_ANALYZE.out.csv + pydamage_filtered_results: PYDAMAGE_FILTER.out.csv + + + +} + From 982503f921e4570b2406ea034c7cdae31d8c4e1a Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 2 Nov 2021 16:08:19 +0100 Subject: [PATCH 002/246] working aDNA workflow + doc --- conf/ancient_dna.config | 21 +++++++ conf/modules.config | 32 +++++++++++ conf/test_ancient_dna.config | 37 ++++++++++++ docs/output.md | 32 +++++++++++ docs/usage.md | 2 + modules/local/bowtie2_assembly_align.nf | 8 ++- .../modules/freebayes/germline/main.nf | 4 +- nextflow.config | 8 +++ nextflow_schema.json | 57 +++++++++++++++---- subworkflows/local/ancient_dna.nf | 56 ++++++++++-------- subworkflows/local/metabat2_binning.nf | 1 + workflows/mag.nf | 22 +++++-- 12 files changed, 237 insertions(+), 43 deletions(-) create mode 100644 conf/ancient_dna.config create mode 100644 conf/test_ancient_dna.config diff --git a/conf/ancient_dna.config b/conf/ancient_dna.config new file mode 100644 index 00000000..bbf42386 --- /dev/null +++ b/conf/ancient_dna.config @@ -0,0 +1,21 @@ +/* +======================================================================================== + Nextflow config file for running minimal tests +======================================================================================== + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run nf-core/mag -profile test, + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Ancient DNA' + config_profile_description = 'Ancient DNA assembly validation profile' + ancient_dna = true + binning_map_mode = 'own' + skip_spades = true + skip_spadeshybrid = true + bowtie2_mode = "--very-sensitive -N 1" +} diff --git a/conf/modules.config b/conf/modules.config index b261f0d1..222b11cd 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -49,6 +49,37 @@ params { publish_dir = "QC_shortreads/fastqc" suffix = ".trimmed" } + 'freebayes' { + args = "-p ${params.freebayes_ploidy}" + publish_dir = "Ancient_DNA/variant_calling/unfiltered" + publish_files = ['vcf.gz': ''] + } + 'bcftools_view' { + args = "-i '%QUAL>=${params.bcftools_view_variant_quality}'" + suffix = ".filtered" + publish_dir = "Ancient_DNA/variant_calling/filtered" + publish_files = ['vcf.gz': ''] + } + 'bcftools_consensus' { + publish_dir = "Ancient_DNA/variant_calling/consensus" + publish_files = ['fa': ''] + } + 'bcftools_index' { + args = "-t" + publish_files = false + } + 'pydamage_analyze' { + publish_dir = "Ancient_DNA/pydamage/analyze" + publish_by_meta = true + } + 'pydamage_filter' { + args = "-t ${params.pydamage_accuracy}" + publish_dir = "Ancient_DNA/pydamage/filter" + publish_by_meta = true + } + 'samtools_faidx' { + publish_files = false + } 'nanolyse' { publish_files = ['log':''] publish_dir = "QC_longreads/NanoLyse" @@ -97,6 +128,7 @@ params { publish_dir = "Assembly" } 'bowtie2_assembly_align' { + args = params.bowtie2_mode publish_files = ['log':''] publish_by_meta = ['assembler', 'QC', 'id'] publish_dir = "Assembly" diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config new file mode 100644 index 00000000..798ad500 --- /dev/null +++ b/conf/test_ancient_dna.config @@ -0,0 +1,37 @@ +/* +======================================================================================== + Nextflow config file for running minimal tests +======================================================================================== + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run nf-core/mag -profile test, + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Ancient DNA test profile ' + config_profile_description = 'Minimal test dataset to check pipeline function for ancient DNA step' + + // Limit resources so that this can run on GitHub Actions + max_cpus = 2 + max_memory = 6.GB + max_time = 6.h + + // Input data + input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' + centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" + kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" + skip_krona = true + min_length_unbinned_contigs = 1 + max_unbinned_contigs = 2 + busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" + gtdb = false + ancient_dna = true + binning_map_mode = 'own' + skip_spades = true + skip_spadeshybrid = true + bcftools_view_variant_quality = 0 + bowtie2_mode = "--very-sensitive -N 1" +} diff --git a/docs/output.md b/docs/output.md index 13afad3b..1b3d9f60 100644 --- a/docs/output.md +++ b/docs/output.md @@ -400,6 +400,38 @@ If the parameters `--cat_db_generate` and `--save_cat_db` are set, additionally +## Ancient DNA (optional, only with `-profile ancient_dna`) + +### `PyDamage` + +[Pydamage](https://github.com/maxibor/pydamage), is tool to automate the process of ancient DNA damage identification and estimation + +
+Output files + +* `Ancient_DNA/pydamage/analyze` + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/latest/output.html](https://pydamage.readthedocs.io/en/latest/output.html) +* `Ancient_DNA/pydamage/filter` + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/latest/output.html](https://pydamage.readthedocs.io/en/latest/output.html) + +
+ +### `variant_calling` + +Because of aDNA damage, *de novo* assemblers sometimes struggle to call a correct consensus on the contig sequence. To avoid this situation, the consensus is re-called with a variant calling software using the reads aligned back to the contigs + +
+Output files + +* `variant_calling/consensus` + * `[sample/group].fa`: contigs sequence with re-called consensus from read-to-contig alignment +* `variant_calling/unfiltered` + * `[sample/group].vcf.gz`: raw variant calls of the reads aligned back to the contigs. +* `variant_calling/filtered` + * `[sample/group].filtered.vcf.gz`: quality filtered variant calls of the reads aligned back to the contigs. + +
+ ### MultiQC
diff --git a/docs/usage.md b/docs/usage.md index a89959c8..1c37eb65 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -148,6 +148,8 @@ If `-profile` is not specified, the pipeline will run locally and expect all sof * `test`, `test_hybrid`, `test_host_rm`, `test_hybrid_host_rm`, `test_busco_auto` * Profiles with a complete configuration for automated testing * Includes links to test data so needs no other parameters +* `ancient_dna` + * A profile to run MAG with ancient DNA assembly specific parameters and the ancient DNA subworkflow. ### `-resume` diff --git a/modules/local/bowtie2_assembly_align.nf b/modules/local/bowtie2_assembly_align.nf index 329472e7..37239cdb 100644 --- a/modules/local/bowtie2_assembly_align.nf +++ b/modules/local/bowtie2_assembly_align.nf @@ -32,7 +32,13 @@ process BOWTIE2_ASSEMBLY_ALIGN { def input = params.single_end ? "-U \"${reads}\"" : "-1 \"${reads[0]}\" -2 \"${reads[1]}\"" """ INDEX=`find -L ./ -name "*.rev.1.bt2l" -o -name "*.rev.1.bt2" | sed 's/.rev.1.bt2l//' | sed 's/.rev.1.bt2//'` - bowtie2 -p "${task.cpus}" -x \$INDEX $input 2> "${name}.bowtie2.log" | \ + bowtie2 \\ + -p "${task.cpus}" \\ + -x \$INDEX \\ + $options.args \\ + $input \\ + 2> "${name}.bowtie2.log" | \ + samtools view -@ "${task.cpus}" -bS | \ samtools sort -@ "${task.cpus}" -o "${name}.bam" samtools index "${name}.bam" diff --git a/modules/nf-core/modules/freebayes/germline/main.nf b/modules/nf-core/modules/freebayes/germline/main.nf index eae62036..4c58dd5a 100644 --- a/modules/nf-core/modules/freebayes/germline/main.nf +++ b/modules/nf-core/modules/freebayes/germline/main.nf @@ -41,7 +41,7 @@ process FREEBAYES_GERMLINE { if (task.cpus > 1) { """ freebayes-parallel \\ - <(fasta_generate_regions.py ${fasta}.fai 10000) ${task.cpus} \\ + <(fasta_generate_regions.py $fai 10000) ${task.cpus} \\ -f $fasta \\ $targets_file \\ $samples_file \\ @@ -50,7 +50,7 @@ process FREEBAYES_GERMLINE { $options.args \\ $input > ${prefix}.vcf - gzip --no-name ${prefix}.vcf + bgzip --threads ${task.cpus} ${prefix}.vcf cat <<-END_VERSIONS > versions.yml ${getProcessName(task.process)}: diff --git a/nextflow.config b/nextflow.config index c27c59b7..8a623fa7 100644 --- a/nextflow.config +++ b/nextflow.config @@ -26,6 +26,7 @@ params { host_removal_save_ids = false // binning options + bowtie2_mode = '' binning_map_mode = 'group' skip_binning = false min_contig_size = 1500 @@ -42,6 +43,12 @@ params { skip_quast = false skip_prodigal = false + // ancient DNA assembly validation options + ancient_dna = false + freebayes_ploidy = 1 + bcftools_view_variant_quality = 10 + pydamage_accuracy = 0.5 + // taxonomy options centrifuge_db = null kraken2_db = null @@ -193,6 +200,7 @@ profiles { test_hybrid_host_rm { includeConfig 'conf/test_hybrid_host_rm.config' } test_busco_auto { includeConfig 'conf/test_busco_auto.config' } test_full { includeConfig 'conf/test_full.config' } + test_ancient_dna { includeConfig 'conf/test_ancient_dna.config' } } // Export these variables to prevent local Python/R libraries from conflicting with those in the container diff --git a/nextflow_schema.json b/nextflow_schema.json index d142199b..f3226435 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -422,33 +422,33 @@ }, "gtdbtk_min_completeness": { "type": "number", - "default": 50.0, + "default": 50, "description": "Min. bin completeness (in %) required to apply GTDB-tk classification.", "help_text": "Completeness assessed with BUSCO analysis (100% - %Missing). Must be greater than 0 (min. 0.01) to avoid GTDB-tk errors. If too low, GTDB-tk classification results can be impaired due to not enough marker genes!", "minimum": 0.01, - "maximum": 100.0 + "maximum": 100 }, "gtdbtk_max_contamination": { "type": "number", - "default": 10.0, + "default": 10, "description": "Max. bin contamination (in %) allowed to apply GTDB-tk classification.", "help_text": "Contamination approximated based on BUSCO analysis (%Complete and duplicated). If too high, GTDB-tk classification results can be impaired due to contamination!", - "minimum": 0.0, - "maximum": 100.0 + "minimum": 0, + "maximum": 100 }, "gtdbtk_min_perc_aa": { "type": "number", "default": 10, "description": "Min. fraction of AA (in %) in the MSA for bins to be kept.", - "minimum": 0.0, - "maximum": 100.0 + "minimum": 0, + "maximum": 100 }, "gtdbtk_min_af": { "type": "number", "default": 0.65, "description": "Min. alignment fraction to consider closest genome.", - "minimum": 0.0, - "maximum": 1.0 + "minimum": 0, + "maximum": 1 }, "gtdbtk_pplacer_cpus": { "type": "number", @@ -547,6 +547,11 @@ "default": 100, "description": "Maximal number of contigs that are not part of any bin but treated as individual genome.", "help_text": "Contigs that do not fulfill the thresholds of `--min_length_unbinned_contigs` and `--max_unbinned_contigs` are pooled for downstream analysis and reporting, except contigs that also do not fullfill `--min_contig_size` are not considered further." + }, + "bowtie2_mode": { + "type": "string", + "description": "Bowtie2 alignment mode", + "help_text": "Bowtie2 alignment mode options, for example: `--very fast` , `--very-sensitive -N 1` , ..." } } }, @@ -580,6 +585,33 @@ "help_text": "Useful to allow reproducibility, as BUSCO datasets are frequently updated and old versions do not always remain accessible." } } + }, + "ancient_dna_assembly": { + "title": "Ancient DNA assembly", + "type": "object", + "description": "", + "default": "", + "properties": { + "ancient_dna": { + "type": "boolean", + "description": "Turn on/off the ancient DNA subworfklow" + }, + "freebayes_ploidy": { + "type": "integer", + "default": 1, + "description": "Ploidy for variant calling" + }, + "bcftools_view_variant_quality": { + "type": "integer", + "default": 10, + "description": "Variant calling quality threshold" + }, + "pydamage_accuracy": { + "type": "number", + "default": 0.5, + "description": "PyDamage accuracy threshold" + } + } } }, "allOf": [ @@ -595,6 +627,9 @@ { "$ref": "#/definitions/max_job_request_options" }, + { + "$ref": "#/definitions/generic_options" + }, { "$ref": "#/definitions/reproducibility_options" }, @@ -620,7 +655,7 @@ "$ref": "#/definitions/bin_quality_check_options" }, { - "$ref": "#/definitions/generic_options" + "$ref": "#/definitions/ancient_dna_assembly" } ] -} +} \ No newline at end of file diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index 7fe14559..f1bd3b89 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -1,35 +1,43 @@ -include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main.nf' addParams( options: [:] ) -include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main.nf' addParams( options: [:] ) -include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main.nf' addParams( options: [:] ) -include { FREEBAYES_GERMLINE as FREEBAYES } from '../../modules/nf-core/modules/freebayes/germline/main.nf' addParams( options: [args : "-p 1"] ) -include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main.nf' addParams( options: [:] ) -include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main.nf' addParams( options: [:] ) -include { SAMTOOLS_FAIDX as FAIDX} from from '../../modules/nf-core/modules/samtools/faidx/main.nf' addParams( options: [:] ) +params.bcftools_consensus_options = [:] +params.bcftools_view_options = [:] +params.bcftools_index_options = [:] +params.freebayes_options = [:] +params.pydamage_analyze_options = [:] +params.pydamage_filter_options = [:] +params.samtools_faidx_options = [:] -worflow ANCIENT_DNA_ASSEMLY_VALIDATION { +include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main' addParams( options: params.bcftools_consensus_options ) +include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main' addParams( options: params.bcftools_index_options ) +include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main' addParams( options: params.bcftools_view_options ) +include { FREEBAYES_GERMLINE as FREEBAYES } from '../../modules/nf-core/modules/freebayes/germline/main' addParams( options: params.freebayes_options ) +include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main' addParams( options: params.pydamage_analyze_options ) +include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main' addParams( options: params.pydamage_filter_options ) +include { SAMTOOLS_FAIDX as FAIDX} from '../../modules/nf-core/modules/samtools/faidx/main' addParams( options: params.samtools_faidx_options ) + +workflow ANCIENT_DNA_ASSEMLY_VALIDATION { take: - contigs // assembled contigs [ val(meta), [contigs] ] - bam // reads mapped back on contigs [ val(meta), [bam] ] - bam_index // bam index [ val(meta), [index] ] + input //channel: [val(meta), path(contigs), path(bam), path(bam_index)] main: - PYDAMAGE_ANALYZE(contigs - .join(bam) - .join(bam_index)) + PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) - FAIDX(contigs.map { it -> [ it[1] ] }) - FREEBAYES (bam.join(bam_index), contigs.map { it -> [ it[1] ] }, FAIDX.out.FAI ) + FAIDX(input.map { item -> [ item[1] ] }) + FREEBAYES (input.map { item -> [item[0], item[2], item[3]] }, input.map { item -> [ item[1] ] }, FAIDX.out.fai, [], [], [], [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) - BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi)) // Add filtering parameters + BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) BCFTOOLS_INDEX_POST(BCFTOOLS_VIEW.out.vcf) BCFTOOLS_CONSENSUS(BCFTOOLS_VIEW.out.vcf .join(BCFTOOLS_INDEX_POST.out.tbi) - .join(contigs)) - emit: - contigs_recalled: BCFTOOLS_CONSENSUS.out.fasta - pydamage_results: PYDAMAGE_ANALYZE.out.csv - pydamage_filtered_results: PYDAMAGE_FILTER.out.csv + .join(input.map { item -> [ item[0], item[1] ] })) - - + ch_versions = Channel.empty() + ch_versions = PYDAMAGE_ANALYZE.out.versions.first() + ch_versions = ch_versions.mix(FAIDX.out.versions.first()) + ch_versions = ch_versions.mix(FREEBAYES.out.versions.first()) + ch_versions = ch_versions.mix(BCFTOOLS_CONSENSUS.out.versions.first()) + emit: + contigs_recalled = BCFTOOLS_CONSENSUS.out.fasta // channel: [ val(meta), path(fasta) ] + pydamage_results = PYDAMAGE_ANALYZE.out.csv // channel: [ val(meta), path(csv) ] + pydamage_filtered_results = PYDAMAGE_FILTER.out.csv // channel: [ val(meta), path(csv) ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 2d8b8783..1e83ecd8 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -77,6 +77,7 @@ workflow METABAT2_BINNING { emit: bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } + assembly_mappings = ch_grouped_mappings bowtie2_version = BOWTIE2_ASSEMBLY_ALIGN.out.version bins = METABAT2.out.bins depths_summary = MAG_DEPTHS_SUMMARY.out.summary diff --git a/workflows/mag.nf b/workflows/mag.nf index 012d85c1..9b7ca2d2 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -90,11 +90,11 @@ include { MULTIQC } from '../modules // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) -include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) -include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) - +include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) +include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) +include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' addParams( bcftools_consensus_options: modules['bcftools_consensus'], bcftools_view_options: modules['bcftools_view'], bcftools_index_options: modules['bcftools_index'], freebayes_options: modules['freebayes'], pydamage_analyze_options: modules['pydamage_analyze'], pydamage_filter_options: modules['pydamage_filter'], samtools_faidx_options: modules['samtools_faidx']) /* ======================================================================================== IMPORT NF-CORE MODULES/SUBWORKFLOWS @@ -568,6 +568,18 @@ workflow MAG { } } + /* + ================================================================================ + Ancient DNA + ================================================================================ + */ + + if (params.ancient_dna){ + METABAT2_BINNING.out.assembly_mappings.view() + ANCIENT_DNA_ASSEMLY_VALIDATION(METABAT2_BINNING.out.assembly_mappings) + ch_software_versions = ch_software_versions.mix(ANCIENT_DNA_ASSEMLY_VALIDATION.out.versions.ifEmpty(null)) + } + // // MODULE: Pipeline reporting // From b786084ceda31ecb9fbc6e135bfd210dbe4ac83e Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 2 Nov 2021 16:23:09 +0100 Subject: [PATCH 003/246] update tool citations --- CITATIONS.md | 10 ++++++++++ 1 file changed, 10 insertions(+) diff --git a/CITATIONS.md b/CITATIONS.md index 6113826c..339e7085 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,6 +10,9 @@ ## Pipeline tools +* [BCFtools]() + > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 + * [Bowtie2](https:/dx.doi.org/10.1038/nmeth.1923) > Langmead, B. and Salzberg, S. L. 2012 Fast gapped-read alignment with Bowtie 2. Nature methods, 9(4), p. 357–359. doi: 10.1038/nmeth.1923. @@ -29,6 +32,9 @@ * [Filtlong](https://github.com/rrwick/Filtlong) +* [Freebayes](https://arxiv.org/abs/1207.3907) + > Garrison E, Marth G. Haplotype-based variant detection from short-read sequencing. arXiv preprint arXiv:1207.3907 [q-bio.GN] 2012 + * [GTDB-Tk](https://doi.org/10.1093/bioinformatics/btz848) > Chaumeil, P. A., Mussig, A. J., Hugenholtz, P., & Parks, D. H. (2020). GTDB-Tk: a toolkit to classify genomes with the Genome Taxonomy Database. Bioinformatics , 36(6), 1925–1927. doi: 10.1093/bioinformatics/btz848. @@ -64,6 +70,10 @@ * [SPAdes](https://doi.org/10.1101/gr.213959.116) > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. +* [PyDamage](https://doi.org/10.7717/peerj.11845) + > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 + + ## Data * [Full-size test data](https://doi.org/10.1038/s41587-019-0191-2) From 41ce2daf9ffca8b31387d2020adca1a1cb5a91ab Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 2 Nov 2021 16:24:42 +0100 Subject: [PATCH 004/246] update bcftools citation --- CITATIONS.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CITATIONS.md b/CITATIONS.md index 339e7085..3fe679a1 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,7 +10,7 @@ ## Pipeline tools -* [BCFtools]() +* [BCFtools](https://doi.org/10.1093/gigascience/giab008) > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 * [Bowtie2](https:/dx.doi.org/10.1038/nmeth.1923) From 03f7048de13401992b28fb396d7eaddda506c5e2 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 2 Nov 2021 16:34:28 +0100 Subject: [PATCH 005/246] update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 089da293..7529412f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,6 +10,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#240](https://github.com/nf-core/mag/pull/240) - Add prodigal to predict protein-coding genes for assemblies. - [#241](https://github.com/nf-core/mag/pull/241) - Add parameter `--skip_prodigal`. - [#244](https://github.com/nf-core/mag/pull/244) - Add pipeline preprint information. +- [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow ### `Changed` From d4633fd839dd247dd4f80e730b168ee392230eb7 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 9 Nov 2021 16:22:24 +0100 Subject: [PATCH 006/246] update freebayes module --- modules.json | 4 ++-- .../modules/freebayes/{germline => }/functions.nf | 0 .../nf-core/modules/freebayes/{germline => }/main.nf | 11 ++++++----- .../nf-core/modules/freebayes/{germline => }/meta.yml | 9 +++++++-- subworkflows/local/ancient_dna.nf | 4 ++-- 5 files changed, 17 insertions(+), 11 deletions(-) rename modules/nf-core/modules/freebayes/{germline => }/functions.nf (100%) rename modules/nf-core/modules/freebayes/{germline => }/main.nf (87%) rename modules/nf-core/modules/freebayes/{germline => }/meta.yml (94%) diff --git a/modules.json b/modules.json index 23933cfe..48159100 100644 --- a/modules.json +++ b/modules.json @@ -18,8 +18,8 @@ "fastqc": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, - "freebayes/germline": { - "git_sha": "3df4fe60851c19ec8d55ef3a978ae851eaccfd68" + "freebayes": { + "git_sha": "9767b081b9d808d7b6671d34d48c70afb436a5f1" }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" diff --git a/modules/nf-core/modules/freebayes/germline/functions.nf b/modules/nf-core/modules/freebayes/functions.nf similarity index 100% rename from modules/nf-core/modules/freebayes/germline/functions.nf rename to modules/nf-core/modules/freebayes/functions.nf diff --git a/modules/nf-core/modules/freebayes/germline/main.nf b/modules/nf-core/modules/freebayes/main.nf similarity index 87% rename from modules/nf-core/modules/freebayes/germline/main.nf rename to modules/nf-core/modules/freebayes/main.nf index 4c58dd5a..0b23dc40 100644 --- a/modules/nf-core/modules/freebayes/germline/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -4,7 +4,7 @@ include { initOptions; saveFiles; getProcessName; getSoftwareName } from './func params.options = [:] options = initOptions(params.options) -process FREEBAYES_GERMLINE { +process FREEBAYES { tag "$meta.id" label 'process_low' publishDir "${params.outdir}", @@ -19,7 +19,7 @@ process FREEBAYES_GERMLINE { } input: - tuple val(meta), path(input), path(input_index) + tuple val(meta), path(input_1), path(input_1_index), path(input_2), path(input_2_index) path fasta path fai path targets @@ -33,6 +33,7 @@ process FREEBAYES_GERMLINE { script: def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def input = input_2 ? "${input_1} ${input_2}" : "${input_1}" def targets_file = targets ? "--target ${targets}" : "" def samples_file = samples ? "--samples ${samples}" : "" def populations_file = populations ? "--populations ${populations}" : "" @@ -41,16 +42,16 @@ process FREEBAYES_GERMLINE { if (task.cpus > 1) { """ freebayes-parallel \\ - <(fasta_generate_regions.py $fai 10000) ${task.cpus} \\ + <(fasta_generate_regions.py ${fasta}.fai 10000) ${task.cpus} \\ -f $fasta \\ $targets_file \\ $samples_file \\ $populations_file \\ $cnv_file \\ $options.args \\ - $input > ${prefix}.vcf + $input > ${prefix}.vcf - bgzip --threads ${task.cpus} ${prefix}.vcf + gzip --no-name ${prefix}.vcf cat <<-END_VERSIONS > versions.yml ${getProcessName(task.process)}: diff --git a/modules/nf-core/modules/freebayes/germline/meta.yml b/modules/nf-core/modules/freebayes/meta.yml similarity index 94% rename from modules/nf-core/modules/freebayes/germline/meta.yml rename to modules/nf-core/modules/freebayes/meta.yml index 86650715..75d44826 100644 --- a/modules/nf-core/modules/freebayes/germline/meta.yml +++ b/modules/nf-core/modules/freebayes/meta.yml @@ -1,11 +1,14 @@ -name: freebayes_germline +name: freebayes description: A haplotype-based variant detector keywords: - variant caller - SNP - genotyping - - variant calling + - somatic variant calling + - germline variant calling + - bacterial variant calling - bayesian + tools: - freebayes: description: Bayesian haplotype-based polymorphism discovery and genotyping @@ -73,6 +76,8 @@ output: type: file description: Compressed VCF file pattern: "*.vcf.gz" + authors: - "@maxibor" - "@FriederikeHanssen" + - "@maxulysse" diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index f1bd3b89..5b8bb5ee 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -9,7 +9,7 @@ params.samtools_faidx_options = [:] include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main' addParams( options: params.bcftools_consensus_options ) include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main' addParams( options: params.bcftools_index_options ) include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main' addParams( options: params.bcftools_view_options ) -include { FREEBAYES_GERMLINE as FREEBAYES } from '../../modules/nf-core/modules/freebayes/germline/main' addParams( options: params.freebayes_options ) +include { FREEBAYES } from '../../modules/nf-core/modules/freebayes/main' addParams( options: params.freebayes_options ) include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main' addParams( options: params.pydamage_analyze_options ) include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main' addParams( options: params.pydamage_filter_options ) include { SAMTOOLS_FAIDX as FAIDX} from '../../modules/nf-core/modules/samtools/faidx/main' addParams( options: params.samtools_faidx_options ) @@ -21,7 +21,7 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) FAIDX(input.map { item -> [ item[1] ] }) - FREEBAYES (input.map { item -> [item[0], item[2], item[3]] }, input.map { item -> [ item[1] ] }, FAIDX.out.fai, [], [], [], [] ) + FREEBAYES (input.map { item -> [item[0], item[2], item[3], [], []] }, input.map { item -> [ item[1] ] }, FAIDX.out.fai, [], [], [], [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) BCFTOOLS_INDEX_POST(BCFTOOLS_VIEW.out.vcf) From 7aa1f039a2cddde94db71937f9e5dc44044c6eb4 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 9 Nov 2021 17:06:05 +0100 Subject: [PATCH 007/246] review comments --- docs/output.md | 6 +++--- docs/usage.md | 6 +++++- workflows/mag.nf | 1 - 3 files changed, 8 insertions(+), 5 deletions(-) diff --git a/docs/output.md b/docs/output.md index 29391077..70d888aa 100644 --- a/docs/output.md +++ b/docs/output.md @@ -430,15 +430,15 @@ Whole genome annotation is the process of identifying features of interest in a ### `PyDamage` -[Pydamage](https://github.com/maxibor/pydamage), is tool to automate the process of ancient DNA damage identification and estimation +[Pydamage](https://github.com/maxibor/pydamage), is a tool to automate the process of ancient DNA damage identification and estimation
Output files * `Ancient_DNA/pydamage/analyze` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/latest/output.html](https://pydamage.readthedocs.io/en/latest/output.html) + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) * `Ancient_DNA/pydamage/filter` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/latest/output.html](https://pydamage.readthedocs.io/en/latest/output.html) + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html)
diff --git a/docs/usage.md b/docs/usage.md index 1c37eb65..4af31795 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -66,6 +66,10 @@ Please note the following requirements: Again, by default, the group information is only used to compute co-abundances for the binning step, but not for group-wise co-assembly (see the parameter docs for [`--coassemble_group`](https://nf-co.re/mag/parameters#coassemble_group) and [`--binning_map_mode`](https://nf-co.re/mag/parameters#binning_map_mode) for more information about how this group information can be used). +### Ancient DNA mode + +Using the ancient DNA profile `-profile ancient_dna` (see [`profile` section](https://nf-co.re/mag/2.1.0/usage#profile)), you can perform ancient DNA *de novo* assembly validation. + ## Running the pipeline The typical command for running the pipeline is as follows: @@ -149,7 +153,7 @@ If `-profile` is not specified, the pipeline will run locally and expect all sof * Profiles with a complete configuration for automated testing * Includes links to test data so needs no other parameters * `ancient_dna` - * A profile to run MAG with ancient DNA assembly specific parameters and the ancient DNA subworkflow. + * A profile to run nf-core/mag with ancient DNA assembly specific parameters and the ancient DNA subworkflow. ### `-resume` diff --git a/workflows/mag.nf b/workflows/mag.nf index 4fc05def..81798a03 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -596,7 +596,6 @@ workflow MAG { */ if (params.ancient_dna){ - METABAT2_BINNING.out.assembly_mappings.view() ANCIENT_DNA_ASSEMLY_VALIDATION(METABAT2_BINNING.out.assembly_mappings) ch_software_versions = ch_software_versions.mix(ANCIENT_DNA_ASSEMLY_VALIDATION.out.versions.ifEmpty(null)) } From 0f7e4f2b9e413c61006812006e79d597da3d6808 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 9 Nov 2021 17:11:15 +0100 Subject: [PATCH 008/246] fix markdownlint --- CITATIONS.md | 5 ++--- docs/output.md | 4 ++-- 2 files changed, 4 insertions(+), 5 deletions(-) diff --git a/CITATIONS.md b/CITATIONS.md index d7677c69..c3dc07c2 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -11,7 +11,7 @@ ## Pipeline tools * [BCFtools](https://doi.org/10.1093/gigascience/giab008) - > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 + > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 * [Bowtie2](https:/dx.doi.org/10.1038/nmeth.1923) > Langmead, B. and Salzberg, S. L. 2012 Fast gapped-read alignment with Bowtie 2. Nature methods, 9(4), p. 357–359. doi: 10.1038/nmeth.1923. @@ -74,8 +74,7 @@ > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. * [PyDamage](https://doi.org/10.7717/peerj.11845) - > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 - + > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 ## Data diff --git a/docs/output.md b/docs/output.md index 70d888aa..70e7458a 100644 --- a/docs/output.md +++ b/docs/output.md @@ -436,9 +436,9 @@ Whole genome annotation is the process of identifying features of interest in a Output files * `Ancient_DNA/pydamage/analyze` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) * `Ancient_DNA/pydamage/filter` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) + * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html)
From 4287f398409afc44a00daecb78883c50d36457e3 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 9 Nov 2021 17:16:37 +0100 Subject: [PATCH 009/246] fix forgotten merge conflict --- nextflow_schema.json | 3 --- 1 file changed, 3 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 3d55f646..99301202 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -657,12 +657,9 @@ }, { "$ref": "#/definitions/bin_quality_check_options" -<<<<<<< HEAD }, { "$ref": "#/definitions/ancient_dna_assembly" -======= ->>>>>>> upstream/dev } ] } \ No newline at end of file From 41b7799459704a01d73e157c8fd7ff66b155f948 Mon Sep 17 00:00:00 2001 From: maxibor Date: Tue, 9 Nov 2021 17:25:08 +0100 Subject: [PATCH 010/246] fix ECLint --- subworkflows/local/ancient_dna.nf | 8 ++++---- subworkflows/local/metabat2_binning.nf | 2 +- workflows/mag.nf | 2 +- 3 files changed, 6 insertions(+), 6 deletions(-) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index 5b8bb5ee..73ad5910 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -23,12 +23,12 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { FAIDX(input.map { item -> [ item[1] ] }) FREEBAYES (input.map { item -> [item[0], item[2], item[3], [], []] }, input.map { item -> [ item[1] ] }, FAIDX.out.fai, [], [], [], [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) - BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) + BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) BCFTOOLS_INDEX_POST(BCFTOOLS_VIEW.out.vcf) BCFTOOLS_CONSENSUS(BCFTOOLS_VIEW.out.vcf - .join(BCFTOOLS_INDEX_POST.out.tbi) - .join(input.map { item -> [ item[0], item[1] ] })) - + .join(BCFTOOLS_INDEX_POST.out.tbi) + .join(input.map { item -> [ item[0], item[1] ] })) + ch_versions = Channel.empty() ch_versions = PYDAMAGE_ANALYZE.out.versions.first() ch_versions = ch_versions.mix(FAIDX.out.versions.first()) diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 1e83ecd8..f10e7dc3 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -77,7 +77,7 @@ workflow METABAT2_BINNING { emit: bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } - assembly_mappings = ch_grouped_mappings + assembly_mappings = ch_grouped_mappings bowtie2_version = BOWTIE2_ASSEMBLY_ALIGN.out.version bins = METABAT2.out.bins depths_summary = MAG_DEPTHS_SUMMARY.out.summary diff --git a/workflows/mag.nf b/workflows/mag.nf index 81798a03..11791c1e 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -94,7 +94,7 @@ include { INPUT_CHECK } from '../subworkflows/local/input_che include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) -include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' addParams( bcftools_consensus_options: modules['bcftools_consensus'], bcftools_view_options: modules['bcftools_view'], bcftools_index_options: modules['bcftools_index'], freebayes_options: modules['freebayes'], pydamage_analyze_options: modules['pydamage_analyze'], pydamage_filter_options: modules['pydamage_filter'], samtools_faidx_options: modules['samtools_faidx']) +include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' addParams( bcftools_consensus_options: modules['bcftools_consensus'], bcftools_view_options: modules['bcftools_view'], bcftools_index_options: modules['bcftools_index'], freebayes_options: modules['freebayes'], pydamage_analyze_options: modules['pydamage_analyze'], pydamage_filter_options: modules['pydamage_filter'], samtools_faidx_options: modules['samtools_faidx']) /* ======================================================================================== IMPORT NF-CORE MODULES/SUBWORKFLOWS From d8e8dc057afc3d190d44895e1ca7796a7cda6061 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 25 Nov 2021 13:16:37 +0100 Subject: [PATCH 011/246] feat: split pre-binning mapping from binning --- subworkflows/local/binning_preparation.nf | 53 +++++++++++++++++++++++ subworkflows/local/metabat2_binning.nf | 44 +++---------------- workflows/mag.nf | 17 ++++++-- 3 files changed, 71 insertions(+), 43 deletions(-) create mode 100644 subworkflows/local/binning_preparation.nf diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf new file mode 100644 index 00000000..eedc12d5 --- /dev/null +++ b/subworkflows/local/binning_preparation.nf @@ -0,0 +1,53 @@ +/* + * Binning preparation with Bowtie2 + */ + +params.bowtie2_build_options = [:] +params.bowtie2_align_options = [:] + +include { BOWTIE2_ASSEMBLY_BUILD } from '../../modules/local/bowtie2_assembly_build' addParams( options: params.bowtie2_build_options ) +include { BOWTIE2_ASSEMBLY_ALIGN } from '../../modules/local/bowtie2_assembly_align' addParams( options: params.bowtie2_align_options ) + +workflow BINNING_PREPARATION { + take: + assemblies // channel: [ val(meta), path(assembly) ] + reads // channel: [ val(meta), [ reads ] ] + + main: + // build bowtie2 index for all assemblies + BOWTIE2_ASSEMBLY_BUILD ( assemblies ) + + // combine assemblies with sample reads for binning depending on specified mapping mode + if (params.binning_map_mode == 'all'){ + // combine assemblies with reads of all samples + ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index + .combine(reads) + } else if (params.binning_map_mode == 'group'){ + // combine assemblies with reads of samples from same group + ch_reads_bowtie2 = reads.map{ meta, reads -> [ meta.group, meta, reads ] } + ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index + .map { meta, assembly, index -> [ meta.group, meta, assembly, index ] } + .combine(ch_reads_bowtie2, by: 0) + .map { group, assembly_meta, assembly, index, reads_meta, reads -> [ assembly_meta, assembly, index, reads_meta, reads ] } + } else { + // combine assemblies (not co-assembled) with reads from own sample + ch_reads_bowtie2 = reads.map{ meta, reads -> [ meta.id, meta, reads ] } + ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index + .map { meta, assembly, index -> [ meta.id, meta, assembly, index ] } + .combine(ch_reads_bowtie2, by: 0) + .map { id, assembly_meta, assembly, index, reads_meta, reads -> [ assembly_meta, assembly, index, reads_meta, reads ] } + } + + BOWTIE2_ASSEMBLY_ALIGN ( ch_bowtie2_input ) + // group mappings for one assembly + ch_grouped_mappings = BOWTIE2_ASSEMBLY_ALIGN.out.mappings + .groupTuple(by: 0) + .map { meta, assembly, bams, bais -> [ meta, assembly[0], bams, bais ] } // multiple symlinks to the same assembly -> use first + + ch_grouped_mappings.dump(tag: "grouped_mappings") + + emit: + bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } + bowtie2_version = BOWTIE2_ASSEMBLY_ALIGN.out.version + grouped_mappings = ch_grouped_mappings +} diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 7e1d03e9..8a583808 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -2,15 +2,11 @@ * Binning with MetaBAT2 */ -params.bowtie2_build_options = [:] -params.bowtie2_align_options = [:] params.metabat2_options = [:] params.mag_depths_options = [:] params.mag_depths_plot_options = [:] params.mag_depths_summary_options = [:] -include { BOWTIE2_ASSEMBLY_BUILD } from '../../modules/local/bowtie2_assembly_build' addParams( options: params.bowtie2_build_options ) -include { BOWTIE2_ASSEMBLY_ALIGN } from '../../modules/local/bowtie2_assembly_align' addParams( options: params.bowtie2_align_options ) include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) @@ -26,41 +22,11 @@ def getColNo(filename) { workflow METABAT2_BINNING { take: - assemblies // channel: [ val(meta), path(assembly) ] + assemblies // channel: [ val(meta), path(assembly), path(bams), path(bais) ] reads // channel: [ val(meta), [ reads ] ] main: - // build bowtie2 index for all assemblies - BOWTIE2_ASSEMBLY_BUILD ( assemblies ) - - // combine assemblies with sample reads for binning depending on specified mapping mode - if (params.binning_map_mode == 'all'){ - // combine assemblies with reads of all samples - ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index - .combine(reads) - } else if (params.binning_map_mode == 'group'){ - // combine assemblies with reads of samples from same group - ch_reads_bowtie2 = reads.map{ meta, reads -> [ meta.group, meta, reads ] } - ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index - .map { meta, assembly, index -> [ meta.group, meta, assembly, index ] } - .combine(ch_reads_bowtie2, by: 0) - .map { group, assembly_meta, assembly, index, reads_meta, reads -> [ assembly_meta, assembly, index, reads_meta, reads ] } - } else { - // combine assemblies (not co-assembled) with reads from own sample - ch_reads_bowtie2 = reads.map{ meta, reads -> [ meta.id, meta, reads ] } - ch_bowtie2_input = BOWTIE2_ASSEMBLY_BUILD.out.assembly_index - .map { meta, assembly, index -> [ meta.id, meta, assembly, index ] } - .combine(ch_reads_bowtie2, by: 0) - .map { id, assembly_meta, assembly, index, reads_meta, reads -> [ assembly_meta, assembly, index, reads_meta, reads ] } - } - - BOWTIE2_ASSEMBLY_ALIGN ( ch_bowtie2_input ) - // group mappings for one assembly - ch_grouped_mappings = BOWTIE2_ASSEMBLY_ALIGN.out.mappings - .groupTuple(by: 0) - .map { meta, assembly, bams, bais -> [ meta, assembly[0], bams, bais ] } // multiple symlinks to the same assembly -> use first - - METABAT2 ( ch_grouped_mappings ) + METABAT2 ( assemblies ) // Compute bin depths for different samples (according to `binning_map_mode`) MAG_DEPTHS ( @@ -68,12 +34,14 @@ workflow METABAT2_BINNING { METABAT2.out.depths ) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) - // create file containg group information for all samples + // create file containing group information for all samples ch_sample_groups = reads .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } + // filter MAG depth files: use only those for plotting that contain depths for > 2 samples ch_mag_depths_plot = MAG_DEPTHS.out.depths .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } + MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() @@ -82,8 +50,6 @@ workflow METABAT2_BINNING { MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) emit: - bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } - bowtie2_version = BOWTIE2_ASSEMBLY_ALIGN.out.version bins = METABAT2.out.bins depths_summary = MAG_DEPTHS_SUMMARY.out.summary metabat2_version = METABAT2.out.version diff --git a/workflows/mag.nf b/workflows/mag.nf index 5f639cca..1acfad3c 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -91,7 +91,8 @@ include { MULTIQC } from '../modules // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) @@ -492,14 +493,22 @@ workflow MAG { ch_bowtie2_assembly_multiqc = Channel.empty() ch_busco_summary = Channel.empty() ch_busco_multiqc = Channel.empty() + + ch_assemblies.dump(tag: "input_into_metabat2") + if (!params.skip_binning){ - METABAT2_BINNING ( + BINNING_PREPARATION ( ch_assemblies, ch_short_reads ) - ch_bowtie2_assembly_multiqc = METABAT2_BINNING.out.bowtie2_assembly_multiqc - ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.bowtie2_version.first().ifEmpty(null)) + + METABAT2_BINNING ( + BINNING_PREPARATION.out.grouped_mappings, + ch_short_reads + ) + ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc + ch_software_versions = ch_software_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first().ifEmpty(null)) ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.metabat2_version.first().ifEmpty(null)) if (!params.skip_busco){ From c49fd8babe0f9a29b5ed07da5c29fc13c9b731d1 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 25 Nov 2021 13:53:27 +0100 Subject: [PATCH 012/246] feat: Separated contig depth generation from metabat2 module --- modules/local/metabat2.nf | 12 +-- .../jgisummarizebamcontigdepths/functions.nf | 78 +++++++++++++++++++ .../jgisummarizebamcontigdepths/main.nf | 44 +++++++++++ .../jgisummarizebamcontigdepths/meta.yml | 50 ++++++++++++ workflows/mag.nf | 3 +- 5 files changed, 180 insertions(+), 7 deletions(-) create mode 100644 modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf create mode 100644 modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf create mode 100644 modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml diff --git a/modules/local/metabat2.nf b/modules/local/metabat2.nf index bbec99fc..fe017ce9 100644 --- a/modules/local/metabat2.nf +++ b/modules/local/metabat2.nf @@ -19,7 +19,7 @@ process METABAT2 { } input: - tuple val(meta), path(assembly), path(bam), path(bai) + tuple val(meta), path(assembly), path(bam), path(bai), path(depth) output: tuple val(meta), path("MetaBAT2/*.fa") , emit: bins @@ -30,11 +30,13 @@ process METABAT2 { script: def software = getSoftwareName(task.process) """ - OMP_NUM_THREADS=${task.cpus} jgi_summarize_bam_contig_depths --outputDepth depth.txt ${bam} - metabat2 -t "${task.cpus}" -i "${assembly}" -a depth.txt -o "MetaBAT2/${meta.assembler}-${meta.id}" -m ${params.min_contig_size} --unbinned --seed ${params.metabat_rng_seed} + ## OMP_NUM_THREADS=${task.cpus} jgi_summarize_bam_contig_depths --outputDepth depth.txt ${bam} + gunzip -f $depth - gzip depth.txt - mv depth.txt.gz "${meta.assembler}-${meta.id}-depth.txt.gz" + metabat2 -t "${task.cpus}" -i "${assembly}" -a ${depth.getName().replace(".gz", "")} -o "MetaBAT2/${meta.assembler}-${meta.id}" -m ${params.min_contig_size} --unbinned --seed ${params.metabat_rng_seed} + + gzip ${depth.getName().replace(".gz", "")} + mv $depth "${meta.assembler}-${meta.id}-depth.txt.gz" # save unbinned contigs above thresholds into individual files, dump others in one file split_fasta.py "MetaBAT2/${meta.assembler}-${meta.id}.unbinned.fa" ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf new file mode 100644 index 00000000..1860ae16 --- /dev/null +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf @@ -0,0 +1,44 @@ +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::metabat2=2.15" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1" + } else { + container "quay.io/biocontainers/metabat2:2.15--h986a166_1" + } + + input: + tuple val(meta), path(bam), path(bai) + + output: + tuple val(meta), path("*.txt.gz"), emit: depth + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + """ + export OMP_NUM_THREADS=$task.cpus + + jgi_summarize_bam_contig_depths \\ + --outputDepth ${prefix}.txt \\ + $options.args \\ + $bam + + bgzip --threads $task.cpus ${prefix}.txt + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml new file mode 100644 index 00000000..351a4701 --- /dev/null +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml @@ -0,0 +1,50 @@ +name: metabat2_jgisummarizebamcontigdepths +description: Depth computation per contig step of metabat2 +keywords: + - sort + - binning + - depth + - bam + - coverage + - de novo assembly +tools: + - metabat2: + description: Metagenome binning + homepage: https://bitbucket.org/berkeleylab/metabat/src/master/ + documentation: https://bitbucket.org/berkeleylab/metabat/src/master/ + tool_dev_url: https://bitbucket.org/berkeleylab/metabat/src/master/ + doi: "10.7717/peerj.7359" + licence: ['BSD-3-clause-LBNL'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: Sorted BAM file of reads aligned on the assembled contigs + pattern: "*.bam" + - bai: + type: file + description: BAM index file + pattern: "*.bam.bai" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - depth: + type: file + description: Text file listing the coverage per contig + pattern: ".txt.gz" + +authors: + - "@maxibor" diff --git a/workflows/mag.nf b/workflows/mag.nf index 1acfad3c..968f244d 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -494,8 +494,6 @@ workflow MAG { ch_busco_summary = Channel.empty() ch_busco_multiqc = Channel.empty() - ch_assemblies.dump(tag: "input_into_metabat2") - if (!params.skip_binning){ BINNING_PREPARATION ( @@ -510,6 +508,7 @@ workflow MAG { ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_software_versions = ch_software_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first().ifEmpty(null)) ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.metabat2_version.first().ifEmpty(null)) + ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first().ifEmpty(null)) if (!params.skip_busco){ /* From f3f4b05f916921a5451bf8be1e7f3fa0b1ab41ba Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 25 Nov 2021 13:58:57 +0100 Subject: [PATCH 013/246] fix: bug in jgisummarisecontigdepths version reporting --- subworkflows/local/metabat2_binning.nf | 32 +++++++++++++++++++------- 1 file changed, 24 insertions(+), 8 deletions(-) diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 8a583808..3f1eb7ff 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -7,10 +7,12 @@ params.mag_depths_options = [:] params.mag_depths_plot_options = [:] params.mag_depths_summary_options = [:] -include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) -include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) -include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) -include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) +include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) // local + +include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' +include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) +include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) +include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) /* * Get number of columns in file (first line) @@ -26,7 +28,20 @@ workflow METABAT2_BINNING { reads // channel: [ val(meta), [ reads ] ] main: - METABAT2 ( assemblies ) + + ch_summarizedepth_input = assemblies + .map { meta, assembly, bams, bais -> [ meta, bams, bais ] } + + METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( + ch_summarizedepth_input + ) + + // TODO re-merge with FASTA assemblies + ch_metabat2_input = assemblies + .join(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth) + .dump(tag: "METABAT2_BINNING:ch_metabat2_input") + + METABAT2 ( ch_metabat2_input ) // Compute bin depths for different samples (according to `binning_map_mode`) MAG_DEPTHS ( @@ -50,7 +65,8 @@ workflow METABAT2_BINNING { MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) emit: - bins = METABAT2.out.bins - depths_summary = MAG_DEPTHS_SUMMARY.out.summary - metabat2_version = METABAT2.out.version + bins = METABAT2.out.bins + depths_summary = MAG_DEPTHS_SUMMARY.out.summary + metabat2_version = METABAT2.out.version + metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions } From 801d216725de5ff7ea898335a7f2cd55ff4e3741 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 25 Nov 2021 15:25:03 +0100 Subject: [PATCH 014/246] fix: partially fix depth naming of now independent jgicontigdepths, add next set of modules --- conf/modules.config | 4 + modules.json | 9 +++ modules/nf-core/modules/maxbin2/functions.nf | 78 ++++++++++++++++++ modules/nf-core/modules/maxbin2/main.nf | 53 +++++++++++++ modules/nf-core/modules/maxbin2/meta.yml | 79 +++++++++++++++++++ .../modules/metabat2/metabat2/functions.nf | 78 ++++++++++++++++++ .../nf-core/modules/metabat2/metabat2/main.nf | 53 +++++++++++++ .../modules/metabat2/metabat2/meta.yml | 56 +++++++++++++ subworkflows/local/metabat2_binning.nf | 12 +-- workflows/mag.nf | 3 +- 10 files changed, 418 insertions(+), 7 deletions(-) create mode 100644 modules/nf-core/modules/maxbin2/functions.nf create mode 100644 modules/nf-core/modules/maxbin2/main.nf create mode 100644 modules/nf-core/modules/maxbin2/meta.yml create mode 100644 modules/nf-core/modules/metabat2/metabat2/functions.nf create mode 100644 modules/nf-core/modules/metabat2/metabat2/main.nf create mode 100644 modules/nf-core/modules/metabat2/metabat2/meta.yml diff --git a/conf/modules.config b/conf/modules.config index c02f1460..61506fe6 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -172,5 +172,9 @@ params { output_format = "gff" publish_by_meta = ['assembler', 'id'] } + 'metabat2_jgisummarizecontigdepths' { + publish_dir = "GenomeBinning" + suffix = "_depth" + } } } diff --git a/modules.json b/modules.json index 16c860ba..e3719dc9 100644 --- a/modules.json +++ b/modules.json @@ -9,6 +9,15 @@ "fastqc": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, + "maxbin2": { + "git_sha": "bd2baa1e7c0f0e3d052e10406d2f86125c51af20" + }, + "metabat2/jgisummarizebamcontigdepths": { + "git_sha": "e0ada7d219e6e3a71b47a8579d1a822a730ed38d" + }, + "metabat2/metabat2": { + "git_sha": "e0ada7d219e6e3a71b47a8579d1a822a730ed38d" + }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" }, diff --git a/modules/nf-core/modules/maxbin2/functions.nf b/modules/nf-core/modules/maxbin2/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/maxbin2/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/maxbin2/main.nf b/modules/nf-core/modules/maxbin2/main.nf new file mode 100644 index 00000000..bcfa9590 --- /dev/null +++ b/modules/nf-core/modules/maxbin2/main.nf @@ -0,0 +1,53 @@ +// Import generic module functions +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process MAXBIN2 { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::maxbin2=2.2.7" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/maxbin2:2.2.7--he1b5a44_2" + } else { + container "quay.io/biocontainers/maxbin2:2.2.7--he1b5a44_2" + } + + input: + tuple val(meta), path(contigs), path(reads), path(abund) + + output: + tuple val(meta), path("*.fasta.gz") , emit: binned_fastas + tuple val(meta), path("*.summary") , emit: summary + tuple val(meta), path("*.log.gz") , emit: log + tuple val(meta), path("*.marker.gz") , emit: marker_counts + tuple val(meta), path("*.noclass.gz") , emit: unbinned_fasta + tuple val(meta), path("*.tooshort.gz"), emit: tooshort_fasta + tuple val(meta), path("*_bin.tar.gz") , emit: marker_bins , optional: true + tuple val(meta), path("*_gene.tar.gz"), emit: marker_genes, optional: true + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def associate_files = reads ? "-reads $reads" : "-abund $abund" + """ + run_MaxBin.pl \\ + -contig $contigs \\ + $associate_files \\ + -thread $task.cpus \\ + $options.args \\ + -out $prefix + + gzip *.fasta *.noclass *.tooshort *log *.marker + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + maxbin2: \$( run_MaxBin.pl -v | head -n 1 | sed 's/MaxBin //' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/maxbin2/meta.yml b/modules/nf-core/modules/maxbin2/meta.yml new file mode 100644 index 00000000..358f8323 --- /dev/null +++ b/modules/nf-core/modules/maxbin2/meta.yml @@ -0,0 +1,79 @@ +name: maxbin2 +description: MaxBin is a software that is capable of clustering metagenomic contigs +keywords: + - metagenomics + - assembly + - binning + - maxbin2 + - de novo assembly + - mags + - metagenome-assembled genomes + - contigs +tools: + - maxbin2: + description: MaxBin is software for binning assembled metagenomic sequences based on an Expectation-Maximization algorithm. + homepage: https://sourceforge.net/projects/maxbin/ + documentation: https://sourceforge.net/projects/maxbin/ + tool_dev_url: https://sourceforge.net/projects/maxbin/ + doi: "10.1093/bioinformatics/btv638" + licence: ['BSD 3-clause'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - contigs: + type: file + description: Multi FASTA file containing assembled contigs of a given sample + pattern: "*.fasta" + - reads: + type: file + description: Reads used to assemble contigs in FASTA or FASTQ format. Do not supply at the same time as abundance files. + pattern: "*.fasta" + - abund: + type: file + description: Contig abundance files, i.e. reads against each contig. See MaxBin2 README for details. Do not supply at the same time as read files. + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - binned_fastas: + type: file + description: Binned contigs, one per bin designated with numeric IDs + pattern: "*.fasta.gz" + - summary: + type: file + description: Summary file describing which contigs are being classified into which bin + pattern: "*.summary" + - log: + type: file + description: Log file recording the core steps of MaxBin algorithm + pattern: "*.log.gz" + - marker: + type: file + description: Marker gene presence numbers for each bin + pattern: "*.marker.gz" + - unbinned_fasta: + type: file + description: All sequences that pass the minimum length threshold but are not classified successfully. + pattern: "*.noclass.gz" + - tooshort_fasta: + type: file + description: All sequences that do not meet the minimum length threshold. + pattern: "*.tooshort.gz" + - marker_genes: + type: file + description: All sequences that do not meet the minimum length threshold. + pattern: "*.marker_of_each_gene.tar.gz" + +authors: + - "@jfy133" diff --git a/modules/nf-core/modules/metabat2/metabat2/functions.nf b/modules/nf-core/modules/metabat2/metabat2/functions.nf new file mode 100644 index 00000000..85628ee0 --- /dev/null +++ b/modules/nf-core/modules/metabat2/metabat2/functions.nf @@ -0,0 +1,78 @@ +// +// Utility functions used in nf-core DSL2 module files +// + +// +// Extract name of software tool from process name using $task.process +// +def getSoftwareName(task_process) { + return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() +} + +// +// Extract name of module from process name using $task.process +// +def getProcessName(task_process) { + return task_process.tokenize(':')[-1] +} + +// +// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules +// +def initOptions(Map args) { + def Map options = [:] + options.args = args.args ?: '' + options.args2 = args.args2 ?: '' + options.args3 = args.args3 ?: '' + options.publish_by_meta = args.publish_by_meta ?: [] + options.publish_dir = args.publish_dir ?: '' + options.publish_files = args.publish_files + options.suffix = args.suffix ?: '' + return options +} + +// +// Tidy up and join elements of a list to return a path string +// +def getPathFromList(path_list) { + def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries + paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes + return paths.join('/') +} + +// +// Function to save/publish module results +// +def saveFiles(Map args) { + def ioptions = initOptions(args.options) + def path_list = [ ioptions.publish_dir ?: args.publish_dir ] + + // Do not publish versions.yml unless running from pytest workflow + if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { + return null + } + if (ioptions.publish_by_meta) { + def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta + for (key in key_list) { + if (args.meta && key instanceof String) { + def path = key + if (args.meta.containsKey(key)) { + path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] + } + path = path instanceof String ? path : '' + path_list.add(path) + } + } + } + if (ioptions.publish_files instanceof Map) { + for (ext in ioptions.publish_files) { + if (args.filename.endsWith(ext.key)) { + def ext_list = path_list.collect() + ext_list.add(ext.value) + return "${getPathFromList(ext_list)}/$args.filename" + } + } + } else if (ioptions.publish_files == null) { + return "${getPathFromList(path_list)}/$args.filename" + } +} diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf new file mode 100644 index 00000000..589e268c --- /dev/null +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -0,0 +1,53 @@ +include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' + +params.options = [:] +options = initOptions(params.options) + +process METABAT2_METABAT2 { + tag "$meta.id" + label 'process_medium' + publishDir "${params.outdir}", + mode: params.publish_dir_mode, + saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } + + conda (params.enable_conda ? "bioconda::metabat2=2.15" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1" + } else { + container "quay.io/biocontainers/metabat2:2.15--h986a166_1" + } + + input: + tuple val(meta), path(fasta), path(depth) + + output: + tuple val(meta), path("bins/*.fa.gz") , optional:true , emit: fasta + tuple val(meta), path("*.tsv.gz"), optional:true , emit: membership + path "versions.yml" , emit: versions + + script: + def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def decompress_depth = depth ? "gzip -d -f $depth" : "" + def depth_file = depth ? "-a ${depth.baseName}" : "" + """ + $decompress_depth + + metabat2 \\ + $options.args \\ + -i $fasta \\ + $depth_file \\ + -t $task.cpus \\ + --saveCls \\ + -o metabat2/${prefix} + + mv metabat2/${prefix} ${prefix}.tsv + mv metabat2 bins + bgzip --threads $task.cpus ${prefix}.tsv + bgzip --threads $task.cpus bins/*.fa + + cat <<-END_VERSIONS > versions.yml + ${getProcessName(task.process)}: + ${getSoftwareName(task.process)}: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/metabat2/metabat2/meta.yml b/modules/nf-core/modules/metabat2/metabat2/meta.yml new file mode 100644 index 00000000..a7f3a7ff --- /dev/null +++ b/modules/nf-core/modules/metabat2/metabat2/meta.yml @@ -0,0 +1,56 @@ +name: metabat2_metabat2 +keywords: + - sort + - binning + - depth + - bam + - coverage + - de novo assembly +tools: + - metabat2: + description: Metagenome binning + homepage: https://bitbucket.org/berkeleylab/metabat/src/master/ + documentation: https://bitbucket.org/berkeleylab/metabat/src/master/ + tool_dev_url: https://bitbucket.org/berkeleylab/metabat/src/master/ + doi: "10.7717/peerj.7359" + licence: ['BSD-3-clause-LBNL'] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Fasta file of the assembled contigs + pattern: "*.{fa,fas,fasta,fna,fa.gz,fas.gz,fasta.gz,fna.gz}" + - depth: + type: file + description: | + Optional text file listing the coverage per contig pre-generated + by metabat2_jgisummarizebamcontigdepths + pattern: "*.txt" + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - fasta: + type: file + description: Bins created from assembled contigs in fasta file + pattern: "*.fa.gz" + - membership: + type: file + description: cluster memberships as a matrix format. + pattern: "*.tsv.gz" + + +authors: + - "@maxibor" diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 3f1eb7ff..af720c25 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -2,14 +2,15 @@ * Binning with MetaBAT2 */ -params.metabat2_options = [:] -params.mag_depths_options = [:] -params.mag_depths_plot_options = [:] -params.mag_depths_summary_options = [:] +params.metabat2_jgisummarizebamcontigdepths_options = [:] +params.metabat2_options = [:] +params.mag_depths_options = [:] +params.mag_depths_plot_options = [:] +params.mag_depths_summary_options = [:] include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) // local -include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' +include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' addParams( options: params.metabat2_jgisummarizebamcontigdepths_options ) include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) @@ -39,7 +40,6 @@ workflow METABAT2_BINNING { // TODO re-merge with FASTA assemblies ch_metabat2_input = assemblies .join(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth) - .dump(tag: "METABAT2_BINNING:ch_metabat2_input") METABAT2 ( ch_metabat2_input ) diff --git a/workflows/mag.nf b/workflows/mag.nf index 968f244d..bbb3d7c6 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -51,6 +51,7 @@ ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multi // Don't overwrite global params.modules, create a copy instead and use that within the main script. def modules = params.modules.clone() def multiqc_options = modules['multiqc'] + multiqc_options.args += params.multiqc_title ? Utils.joinModuleArgs(["--title \"$params.multiqc_title\""]) : '' // @@ -92,7 +93,7 @@ include { MULTIQC } from '../modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) From 3620b7144a276a051fc8e868b74d1d771c3b710a Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 25 Nov 2021 17:45:27 +0100 Subject: [PATCH 015/246] fix: trying Mahesh's suggestion --- subworkflows/local/metabat2_binning.nf | 2 +- workflows/mag.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index af720c25..c1a7fc3c 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -10,7 +10,7 @@ params.mag_depths_summary_options = [:] include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) // local -include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' addParams( options: params.metabat2_jgisummarizebamcontigdepths_options ) +include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' addParams( options: [ suffix: { meta.assembler } ] ) include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) diff --git a/workflows/mag.nf b/workflows/mag.nf index bbb3d7c6..d5c75013 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -93,7 +93,7 @@ include { MULTIQC } from '../modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' //addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) From 2b37ec2584e5f84b0d30cd06f8f804f700cb2730 Mon Sep 17 00:00:00 2001 From: d4straub Date: Fri, 26 Nov 2021 12:05:51 +0100 Subject: [PATCH 016/246] bump version to 2.2.0dev --- CHANGELOG.md | 8 ++++++++ nextflow.config | 2 +- 2 files changed, 9 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 14c488f1..9fe18e26 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,14 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). +## v2.2.0dev + +### `Added` + +### `Changed` + +### `Fixed` + ## v2.1.1 - 2021/11/25 ### `Added` diff --git a/nextflow.config b/nextflow.config index a49cfb7f..04b7cca1 100644 --- a/nextflow.config +++ b/nextflow.config @@ -231,7 +231,7 @@ manifest { description = 'Assembly, binning and annotation of metagenomes' mainScript = 'main.nf' nextflowVersion = '!>=21.04.0' - version = '2.1.1' + version = '2.2.0dev' } // Function to ensure that resource requirements don't go beyond From b052de324d66d337fb29b3b81fe14e993d4c5f11 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 29 Nov 2021 11:33:57 +0100 Subject: [PATCH 017/246] Update modules to new syntax and get depth name working --- conf/modules.config | 24 +++++- docs/output.md | 4 +- modules.json | 6 +- modules/nf-core/modules/maxbin2/functions.nf | 78 ------------------- modules/nf-core/modules/maxbin2/main.nf | 24 ++---- .../jgisummarizebamcontigdepths/functions.nf | 78 ------------------- .../jgisummarizebamcontigdepths/main.nf | 25 ++---- .../modules/metabat2/metabat2/functions.nf | 78 ------------------- .../nf-core/modules/metabat2/metabat2/main.nf | 25 ++---- 9 files changed, 48 insertions(+), 294 deletions(-) delete mode 100644 modules/nf-core/modules/maxbin2/functions.nf delete mode 100644 modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf delete mode 100644 modules/nf-core/modules/metabat2/metabat2/functions.nf diff --git a/conf/modules.config b/conf/modules.config index 61506fe6..58a991d9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -172,9 +172,25 @@ params { output_format = "gff" publish_by_meta = ['assembler', 'id'] } - 'metabat2_jgisummarizecontigdepths' { - publish_dir = "GenomeBinning" - suffix = "_depth" - } } } + +// nf-core modules DSL2 syntax v2.0 +process { + + publishDir = [ + path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + + withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { + publishDir = [ + path: { "${params.outdir}/GenomeBinning/" }, + mode: 'copy', + pattern: '*_depth.txt.gz' + ] + ext.suffix = { "_${meta.assembler}_depth" } + } + +} diff --git a/docs/output.md b/docs/output.md index 644efb1d..3e77ca31 100644 --- a/docs/output.md +++ b/docs/output.md @@ -235,13 +235,13 @@ Protein-coding genes are predicted for each assembly. ### Contig sequencing depth -Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig_depths --outputDepth`. The values correspond to `(sum of exactely aligned bases) / ((contig length)-2*75)`. For example, for two reads aligned exactly with `10` and `9` bases on a 1000 bp long contig the depth is calculated by `(10+9)/(1000-2*75)` (1000bp length of contig minus 75bp from each end, which is excluded). +Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig_depths --outputDepth`. The values correspond to `(sum of exactly aligned bases) / ((contig length)-2*75)`. For example, for two reads aligned exactly with `10` and `9` bases on a 1000 bp long contig the depth is calculated by `(10+9)/(1000-2*75)` (1000bp length of contig minus 75bp from each end, which is excluded).
Output files * `GenomeBinning/` - * `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads. + * `[sample/group]-[assembler]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads.
diff --git a/modules.json b/modules.json index e3719dc9..275f82cf 100644 --- a/modules.json +++ b/modules.json @@ -10,13 +10,13 @@ "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, "maxbin2": { - "git_sha": "bd2baa1e7c0f0e3d052e10406d2f86125c51af20" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "metabat2/jgisummarizebamcontigdepths": { - "git_sha": "e0ada7d219e6e3a71b47a8579d1a822a730ed38d" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "metabat2/metabat2": { - "git_sha": "e0ada7d219e6e3a71b47a8579d1a822a730ed38d" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" diff --git a/modules/nf-core/modules/maxbin2/functions.nf b/modules/nf-core/modules/maxbin2/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/maxbin2/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/maxbin2/main.nf b/modules/nf-core/modules/maxbin2/main.nf index bcfa9590..e13af704 100644 --- a/modules/nf-core/modules/maxbin2/main.nf +++ b/modules/nf-core/modules/maxbin2/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process MAXBIN2 { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::maxbin2=2.2.7" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/maxbin2:2.2.7--he1b5a44_2" - } else { - container "quay.io/biocontainers/maxbin2:2.2.7--he1b5a44_2" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/maxbin2:2.2.7--he1b5a44_2' : + 'quay.io/biocontainers/maxbin2:2.2.7--he1b5a44_2' }" input: tuple val(meta), path(contigs), path(reads), path(abund) @@ -33,20 +22,21 @@ process MAXBIN2 { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" def associate_files = reads ? "-reads $reads" : "-abund $abund" """ run_MaxBin.pl \\ -contig $contigs \\ $associate_files \\ -thread $task.cpus \\ - $options.args \\ + $args \\ -out $prefix gzip *.fasta *.noclass *.tooshort *log *.marker cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: + "${task.process}": maxbin2: \$( run_MaxBin.pl -v | head -n 1 | sed 's/MaxBin //' ) END_VERSIONS """ diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf index 1860ae16..e35d6715 100644 --- a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf @@ -1,21 +1,11 @@ -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::metabat2=2.15" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1" - } else { - container "quay.io/biocontainers/metabat2:2.15--h986a166_1" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1' : + 'quay.io/biocontainers/metabat2:2.15--h986a166_1' }" input: tuple val(meta), path(bam), path(bai) @@ -25,20 +15,21 @@ process METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" """ export OMP_NUM_THREADS=$task.cpus jgi_summarize_bam_contig_depths \\ --outputDepth ${prefix}.txt \\ - $options.args \\ + $args \\ $bam bgzip --threads $task.cpus ${prefix}.txt cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) + "${task.process}": + metabat2: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) END_VERSIONS """ } diff --git a/modules/nf-core/modules/metabat2/metabat2/functions.nf b/modules/nf-core/modules/metabat2/metabat2/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/metabat2/metabat2/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index 589e268c..d158af91 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -1,21 +1,11 @@ -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process METABAT2_METABAT2 { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::metabat2=2.15" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1" - } else { - container "quay.io/biocontainers/metabat2:2.15--h986a166_1" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/metabat2:2.15--h986a166_1' : + 'quay.io/biocontainers/metabat2:2.15--h986a166_1' }" input: tuple val(meta), path(fasta), path(depth) @@ -26,14 +16,15 @@ process METABAT2_METABAT2 { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" def decompress_depth = depth ? "gzip -d -f $depth" : "" def depth_file = depth ? "-a ${depth.baseName}" : "" """ $decompress_depth metabat2 \\ - $options.args \\ + $args \\ -i $fasta \\ $depth_file \\ -t $task.cpus \\ @@ -46,8 +37,8 @@ process METABAT2_METABAT2 { bgzip --threads $task.cpus bins/*.fa cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) + "${task.process}": + metabat2: \$( metabat2 --help 2>&1 | head -n 2 | tail -n 1| sed 's/.*\\:\\([0-9]*\\.[0-9]*\\).*/\\1/' ) END_VERSIONS """ } From 04bad5e39d5177ed1b0df27902d408625872c1f9 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 29 Nov 2021 11:54:35 +0100 Subject: [PATCH 018/246] Update split_fasta to support gzipped input --- bin/split_fasta.py | 54 ++++++++++++++++++++++++++++++---------------- 1 file changed, 36 insertions(+), 18 deletions(-) diff --git a/bin/split_fasta.py b/bin/split_fasta.py index e009e1eb..c7c6e6bc 100755 --- a/bin/split_fasta.py +++ b/bin/split_fasta.py @@ -1,8 +1,9 @@ #!/usr/bin/env python -#USAGE: ./combine_tables.py <*.unbinned.fa> +#USAGE: ./split_fasta.py <*.unbinned.fa(.gz)> import pandas as pd +import gzip from sys import argv from Bio import SeqIO from Bio.Seq import Seq @@ -24,23 +25,40 @@ pooled = [] remaining = [] -# Read file -with open(input_file) as f: - fasta_sequences = SeqIO.parse(f,'fasta') - - for fasta in fasta_sequences: - name, sequence = fasta.id, str(fasta.seq) - length = len(sequence) - - # store each sequence above threshold together with its length into df - if length >= length_threshold: - df_above_threshold = df_above_threshold.append({"id":name, "seq":sequence, "length":length}, ignore_index = True) - # contigs to retain and pool - elif length >= min_length_to_retain_contig: - pooled.append(SeqRecord(Seq(sequence, generic_dna), id = name)) - # remaining sequences - else: - remaining.append(SeqRecord(Seq(sequence, generic_dna), id = name)) +if input_file.endswith('.gz'): + with gzip.open(input_file, 'rt') as f: + fasta_sequences = SeqIO.parse(f,'fasta') + + for fasta in fasta_sequences: + name, sequence = fasta.id, str(fasta.seq) + length = len(sequence) + + # store each sequence above threshold together with its length into df + if length >= length_threshold: + df_above_threshold = df_above_threshold.append({"id":name, "seq":sequence, "length":length}, ignore_index = True) + # contigs to retain and pool + elif length >= min_length_to_retain_contig: + pooled.append(SeqRecord(Seq(sequence, generic_dna), id = name)) + # remaining sequences + else: + remaining.append(SeqRecord(Seq(sequence, generic_dna), id = name)) +else: + with open(input_file) as f: + fasta_sequences = SeqIO.parse(f,'fasta') + + for fasta in fasta_sequences: + name, sequence = fasta.id, str(fasta.seq) + length = len(sequence) + + # store each sequence above threshold together with its length into df + if length >= length_threshold: + df_above_threshold = df_above_threshold.append({"id":name, "seq":sequence, "length":length}, ignore_index = True) + # contigs to retain and pool + elif length >= min_length_to_retain_contig: + pooled.append(SeqRecord(Seq(sequence, generic_dna), id = name)) + # remaining sequences + else: + remaining.append(SeqRecord(Seq(sequence, generic_dna), id = name)) # Sort sequences above threshold by length df_above_threshold.sort_values(by=['length'], ascending=False, inplace=True) From a3c6718df933e6aca992e05a9bba2ace3eb56dda Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 29 Nov 2021 17:14:54 +0100 Subject: [PATCH 019/246] feat: seperated split_fastq and use nf-core metabat. allow get_mag_depths.py to accept gzipped fasta. Stuck on gunzip bins fr downstream --- bin/get_mag_depths.py | 19 +++++++--- conf/modules.config | 16 ++++++-- modules.json | 3 ++ modules/local/mag_depths.nf | 5 +-- modules/local/split_fastq.nf | 28 ++++++++++++++ modules/nf-core/modules/gunzip/main.nf | 31 +++++++++++++++ modules/nf-core/modules/gunzip/meta.yml | 34 +++++++++++++++++ .../nf-core/modules/metabat2/metabat2/main.nf | 11 ++++-- nf-core/modules/gunzip/main.nf | 32 ++++++++++++++++ nf-core/modules/gunzip/meta.yml | 34 +++++++++++++++++ subworkflows/local/binning_preparation.nf | 2 +- subworkflows/local/metabat2_binning.nf | 38 +++++++++++++------ 12 files changed, 226 insertions(+), 27 deletions(-) create mode 100644 modules/local/split_fastq.nf create mode 100644 modules/nf-core/modules/gunzip/main.nf create mode 100644 modules/nf-core/modules/gunzip/meta.yml create mode 100644 nf-core/modules/gunzip/main.nf create mode 100644 nf-core/modules/gunzip/meta.yml diff --git a/bin/get_mag_depths.py b/bin/get_mag_depths.py index 2ade6ec6..6cc547dd 100755 --- a/bin/get_mag_depths.py +++ b/bin/get_mag_depths.py @@ -44,13 +44,22 @@ def main(args=None): n_samples = len(sample_names) # for each bin, access contig depths and compute mean bin depth (for all samples) print("bin", '\t'.join(sample_names), sep='\t', file=args.out) + for file in args.bins: all_depths = [[] for i in range(n_samples)] - with open(file, "rt") as infile: - for rec in SeqIO.parse(infile,'fasta'): - contig_depths = dict_contig_depths[rec.id] - for sample in range(n_samples): - all_depths[sample].append(contig_depths[sample]) + + if file.endswith('.gz'): + with gzip.open(file, 'rt') as infile: + for rec in SeqIO.parse(infile,'fasta'): + contig_depths = dict_contig_depths[rec.id] + for sample in range(n_samples): + all_depths[sample].append(contig_depths[sample]) + else: + with open(file, "rt") as infile: + for rec in SeqIO.parse(infile,'fasta'): + contig_depths = dict_contig_depths[rec.id] + for sample in range(n_samples): + all_depths[sample].append(contig_depths[sample]) print(os.path.basename(file), '\t'.join(str(statistics.median(sample_depths)) for sample_depths in all_depths), sep='\t', file=args.out) diff --git a/conf/modules.config b/conf/modules.config index 58a991d9..c75819d9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -101,10 +101,6 @@ params { publish_by_meta = ['assembler', 'QC', 'id'] publish_dir = "Assembly" } - 'metabat2' { - publish_files = ['txt.gz':'', 'fa':'', 'fa.gz':''] - publish_dir = "GenomeBinning" - } 'mag_depths' { publish_files = false } @@ -193,4 +189,16 @@ process { ext.suffix = { "_${meta.assembler}_depth" } } + withName: METABAT2_METABAT2 { + publishDir = [ + path: { "${params.outdir}/GenomeBinning/" }, + mode: 'copy' + ] + ext.args = [ + "-m ${params.min_contig_size}", + "--unbinned", + "--seed ${params.metabat_rng_seed}" + ].join(' ').trim() + } + } diff --git a/modules.json b/modules.json index 275f82cf..59ae54e2 100644 --- a/modules.json +++ b/modules.json @@ -9,6 +9,9 @@ "fastqc": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, + "gunzip": { + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + }, "maxbin2": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, diff --git a/modules/local/mag_depths.nf b/modules/local/mag_depths.nf index 4b333665..533496b5 100644 --- a/modules/local/mag_depths.nf +++ b/modules/local/mag_depths.nf @@ -20,8 +20,7 @@ process MAG_DEPTHS { } input: - tuple val(meta), path(bins) - path(contig_depths) + tuple val(meta), path(bins), path(contig_depths) output: tuple val(meta), path("${meta.assembler}-${meta.id}-binDepths.tsv"), emit: depths @@ -29,7 +28,7 @@ process MAG_DEPTHS { script: def software = getSoftwareName(task.process) """ - get_mag_depths.py --bins ${bins} \ + get_mag_depths.py --bins *.fa.gz \ --depths ${contig_depths} \ --assembly_name "${meta.assembler}-${meta.id}" \ --out "${meta.assembler}-${meta.id}-binDepths.tsv" diff --git a/modules/local/split_fastq.nf b/modules/local/split_fastq.nf new file mode 100644 index 00000000..2d1d047f --- /dev/null +++ b/modules/local/split_fastq.nf @@ -0,0 +1,28 @@ +process SPLIT_FASTQ { + tag "${meta.assembler}-${meta.id}" + label 'process_low' + + // Using container from metabat2 process, since this will be anyway already downloaded and contains biopython and pandas + conda (params.enable_conda ? "bioconda::metabat2=2.15 conda-forge::python=3.6.7 conda-forge::biopython=1.74 conda-forge::pandas=1.1.5" : null) + if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { + container "https://depot.galaxyproject.org/singularity/mulled-v2-e25d1fa2bb6cbacd47a4f8b2308bd01ba38c5dd7:75310f02364a762e6ba5206fcd11d7529534ed6e-0" + } else { + container "quay.io/biocontainers/mulled-v2-e25d1fa2bb6cbacd47a4f8b2308bd01ba38c5dd7:75310f02364a762e6ba5206fcd11d7529534ed6e-0" + } + + input: + tuple val(meta), path(unbinned) + + output: + path "${meta.assembler}/unbinned/" , emit: unbinned + + script: + """ + # save unbinned contigs above thresholds into individual files, dump others in one file + split_fasta.py $unbinned ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} + + mkdir -p ${meta.assembler}/unbinned/ + mv *.fa ${meta.assembler}/unbinned/ + gzip ${meta.assembler}/unbinned/* + """ +} diff --git a/modules/nf-core/modules/gunzip/main.nf b/modules/nf-core/modules/gunzip/main.nf new file mode 100644 index 00000000..77a4e546 --- /dev/null +++ b/modules/nf-core/modules/gunzip/main.nf @@ -0,0 +1,31 @@ +process GUNZIP { + tag "$archive" + label 'process_low' + + conda (params.enable_conda ? "conda-forge::sed=4.7" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : + 'biocontainers/biocontainers:v1.2.0_cv1' }" + + input: + tuple val(meta), path(archive) + + output: + tuple val(meta), path("$gunzip"), emit: gunzip + path "versions.yml" , emit: versions + + script: + def args = task.ext.args ?: '' + gunzip = archive.toString() - '.gz' + """ + gunzip \\ + -f \\ + $args \\ + $archive + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gunzip: \$(echo \$(gunzip --version 2>&1) | sed 's/^.*(gzip) //; s/ Copyright.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/gunzip/meta.yml b/modules/nf-core/modules/gunzip/meta.yml new file mode 100644 index 00000000..ea1f1546 --- /dev/null +++ b/modules/nf-core/modules/gunzip/meta.yml @@ -0,0 +1,34 @@ +name: gunzip +description: Compresses and decompresses files. +keywords: + - gunzip + - compression +tools: + - gunzip: + description: | + gzip is a file format and a software application used for file compression and decompression. + documentation: https://www.gnu.org/software/gzip/manual/gzip.html + licence: ['GPL-3.0-or-later'] +input: + - meta: + type: map + description: | + Optional groovy Map containing meta information + e.g. [ id:'test', single_end:false ] + - archive: + type: file + description: File to be compressed/uncompressed + pattern: "*.*" +output: + - gunzip: + type: file + description: Compressed/uncompressed file + pattern: "*.*" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" + - "@jfy133" diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index d158af91..1b180fc8 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -12,7 +12,10 @@ process METABAT2_METABAT2 { output: tuple val(meta), path("bins/*.fa.gz") , optional:true , emit: fasta - tuple val(meta), path("*.tsv.gz"), optional:true , emit: membership + tuple val(meta), path("*.tooShort.fa.gz") , optional:true , emit: tooshort + tuple val(meta), path("*.lowDepth.fa.gz") , optional:true , emit: lowdepth + tuple val(meta), path("*.unbinned.fa.gz") , optional:true , emit: unbinned + tuple val(meta), path("*.tsv.gz") , optional:true , emit: membership path "versions.yml" , emit: versions script: @@ -33,8 +36,10 @@ process METABAT2_METABAT2 { mv metabat2/${prefix} ${prefix}.tsv mv metabat2 bins - bgzip --threads $task.cpus ${prefix}.tsv - bgzip --threads $task.cpus bins/*.fa + + gzip ${prefix}.tsv + gzip bins/*.fa + mv bins/*.{tooShort,lowDepth,unbinned}.fa.gz . cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/nf-core/modules/gunzip/main.nf b/nf-core/modules/gunzip/main.nf new file mode 100644 index 00000000..ba9aae48 --- /dev/null +++ b/nf-core/modules/gunzip/main.nf @@ -0,0 +1,32 @@ +process GUNZIP { + tag "$archive" + label 'process_low' + + conda (params.enable_conda ? "conda-forge::sed=4.7" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : + 'biocontainers/biocontainers:v1.2.0_cv1' }" + + input: + tuple val(meta), path(archive) + + output: + tuple val(meta), path("$gunzip"), emit: gunzip + path "versions.yml" , emit: versions + + script: + def args = task.ext.args ?: '' + gunzip = archive.collect { it.toString() - '.gz' } + """ + echo "${archive.toString() - '.gz'}" + gunzip \\ + -f \\ + $args \\ + $archive + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gunzip: \$(echo \$(gunzip --version 2>&1) | sed 's/^.*(gzip) //; s/ Copyright.*\$//') + END_VERSIONS + """ +} diff --git a/nf-core/modules/gunzip/meta.yml b/nf-core/modules/gunzip/meta.yml new file mode 100644 index 00000000..ea1f1546 --- /dev/null +++ b/nf-core/modules/gunzip/meta.yml @@ -0,0 +1,34 @@ +name: gunzip +description: Compresses and decompresses files. +keywords: + - gunzip + - compression +tools: + - gunzip: + description: | + gzip is a file format and a software application used for file compression and decompression. + documentation: https://www.gnu.org/software/gzip/manual/gzip.html + licence: ['GPL-3.0-or-later'] +input: + - meta: + type: map + description: | + Optional groovy Map containing meta information + e.g. [ id:'test', single_end:false ] + - archive: + type: file + description: File to be compressed/uncompressed + pattern: "*.*" +output: + - gunzip: + type: file + description: Compressed/uncompressed file + pattern: "*.*" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" + - "@jfy133" diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf index eedc12d5..96dbf833 100644 --- a/subworkflows/local/binning_preparation.nf +++ b/subworkflows/local/binning_preparation.nf @@ -44,7 +44,7 @@ workflow BINNING_PREPARATION { .groupTuple(by: 0) .map { meta, assembly, bams, bais -> [ meta, assembly[0], bams, bais ] } // multiple symlinks to the same assembly -> use first - ch_grouped_mappings.dump(tag: "grouped_mappings") + ch_grouped_mappings emit: bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index c1a7fc3c..41feb846 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -2,15 +2,15 @@ * Binning with MetaBAT2 */ -params.metabat2_jgisummarizebamcontigdepths_options = [:] -params.metabat2_options = [:] params.mag_depths_options = [:] params.mag_depths_plot_options = [:] params.mag_depths_summary_options = [:] -include { METABAT2 } from '../../modules/local/metabat2' addParams( options: params.metabat2_options ) // local +include { METABAT2_METABAT2 } from '../../modules/nf-core/modules/metabat2/metabat2/main' +include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' +include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' -include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' addParams( options: [ suffix: { meta.assembler } ] ) +include { SPLIT_FASTQ } from '../../modules/local/split_fastq' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) @@ -37,16 +37,29 @@ workflow METABAT2_BINNING { ch_summarizedepth_input ) - // TODO re-merge with FASTA assemblies ch_metabat2_input = assemblies - .join(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth) + .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + . map { it -> [ it[0], it[1], it[4] ] } - METABAT2 ( ch_metabat2_input ) + METABAT2_METABAT2 ( ch_metabat2_input ) + + // split FASTQ + SPLIT_FASTQ ( + METABAT2_METABAT2.out.unbinned + ) + + // decompress main bins for downstream + GUNZIP ( + METABAT2_METABAT2.out.fasta + ) // Compute bin depths for different samples (according to `binning_map_mode`) + ch_depth_input = METABAT2_METABAT2.out.fasta + .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + .dump(tag:"pre_for_mag_depths") + MAG_DEPTHS ( - METABAT2.out.bins, - METABAT2.out.depths + ch_depth_input ) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples @@ -65,8 +78,11 @@ workflow METABAT2_BINNING { MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) emit: - bins = METABAT2.out.bins + bins = GUNZIP.out.gunzip // TODO this would include discarded FASTAS! need to separate out somehow! Probably in metabat2 module + unbinned = SPLIT_FASTQ.out.unbinned + tooshort = METABAT2_METABAT2.out.tooshort + lowdepth = METABAT2_METABAT2.out.lowdepth depths_summary = MAG_DEPTHS_SUMMARY.out.summary - metabat2_version = METABAT2.out.version + metabat2_version = METABAT2_METABAT2.out.versions metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions } From cb778fbb83a8324b2694cec49c0a393aed829ca5 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 2 Dec 2021 15:26:43 +0100 Subject: [PATCH 020/246] Updated modules but metabat2 still broken --- modules.json | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/modules.json b/modules.json index 59ae54e2..dad8c6eb 100644 --- a/modules.json +++ b/modules.json @@ -13,13 +13,13 @@ "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "maxbin2": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "metabat2/jgisummarizebamcontigdepths": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "metabat2/metabat2": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" From 23e74f77aea4fe0c1a6fcd329b6b84967eff3b90 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 3 Dec 2021 14:31:38 +0100 Subject: [PATCH 021/246] update to nf-core DSL2 syntax v2 --- conf/modules.config | 92 ++++++++++++------- modules.json | 20 ++-- .../modules/bcftools/consensus/functions.nf | 78 ---------------- .../modules/bcftools/consensus/main.nf | 26 ++---- .../modules/bcftools/index/functions.nf | 78 ---------------- .../nf-core/modules/bcftools/index/main.nf | 26 ++---- .../modules/bcftools/view/functions.nf | 78 ---------------- modules/nf-core/modules/bcftools/view/main.nf | 28 ++---- .../nf-core/modules/freebayes/functions.nf | 78 ---------------- modules/nf-core/modules/freebayes/main.nf | 40 +++----- modules/nf-core/modules/freebayes/meta.yml | 7 +- .../modules/pydamage/analyze/functions.nf | 78 ---------------- .../nf-core/modules/pydamage/analyze/main.nf | 26 ++---- .../modules/pydamage/filter/functions.nf | 78 ---------------- .../nf-core/modules/pydamage/filter/main.nf | 26 ++---- .../modules/samtools/faidx/functions.nf | 78 ---------------- .../nf-core/modules/samtools/faidx/main.nf | 24 ++--- 17 files changed, 136 insertions(+), 725 deletions(-) delete mode 100644 modules/nf-core/modules/bcftools/consensus/functions.nf delete mode 100644 modules/nf-core/modules/bcftools/index/functions.nf delete mode 100644 modules/nf-core/modules/bcftools/view/functions.nf delete mode 100644 modules/nf-core/modules/freebayes/functions.nf delete mode 100644 modules/nf-core/modules/pydamage/analyze/functions.nf delete mode 100644 modules/nf-core/modules/pydamage/filter/functions.nf delete mode 100644 modules/nf-core/modules/samtools/faidx/functions.nf diff --git a/conf/modules.config b/conf/modules.config index 06998f70..393b0ae1 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -49,37 +49,6 @@ params { publish_dir = "QC_shortreads/fastqc" suffix = ".trimmed" } - 'freebayes' { - args = "-p ${params.freebayes_ploidy}" - publish_dir = "Ancient_DNA/variant_calling/unfiltered" - publish_files = ['vcf.gz': ''] - } - 'bcftools_view' { - args = "-i '%QUAL>=${params.bcftools_view_variant_quality}'" - suffix = ".filtered" - publish_dir = "Ancient_DNA/variant_calling/filtered" - publish_files = ['vcf.gz': ''] - } - 'bcftools_consensus' { - publish_dir = "Ancient_DNA/variant_calling/consensus" - publish_files = ['fa': ''] - } - 'bcftools_index' { - args = "-t" - publish_files = false - } - 'pydamage_analyze' { - publish_dir = "Ancient_DNA/pydamage/analyze" - publish_by_meta = true - } - 'pydamage_filter' { - args = "-t ${params.pydamage_accuracy}" - publish_dir = "Ancient_DNA/pydamage/filter" - publish_by_meta = true - } - 'samtools_faidx' { - publish_files = false - } 'nanolyse' { publish_files = ['log':''] publish_dir = "QC_longreads/NanoLyse" @@ -206,3 +175,64 @@ params { } } } + +process { + + publishDir = [ + path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + + withName: freebayes { + ext.args = "-p ${params.freebayes_ploidy}" + publishDir = [ + path: { "${params.outdir}/Ancient_DNA/variant_calling/freebayes" }, + pattern: "*.vcf.gz" + ] + } + + withName: bcftools_view { + ext.args = "-i '%QUAL>=${params.bcftools_view_variant_quality}'" + ext.suffix = ".filtered" + publishDir = [ + path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, + pattern: "*.vcf.gz" + ] + } + + withName: bcftools_consensus { + publishDir = [ + path: {"${params.outdir}/Ancient_DNA/variant_calling/consensus" }, + pattern: "*.fa" + ] + } + + withName: bcftools_index { + ext.args = "-t" + publishDir = [ + path: {"${params.outdir}/Ancient_DNA/variant_calling/index" }, + enabled: false + ] + } + + withName: pydamage_analyze { + publishDir = [ + path: {"${params.outdir}/Ancient_DNA/pydamage/analyze" } + ] + } + + withName: pydamage_filter { + ext.args = "-t ${params.pydamage_accuracy}" + publishDir = [ + path: {"${params.outdir}/Ancient_DNA/pydamage/filter" } + ] + } + + withName: samtools_faidx { + publishDir = [ + path: {"${params.outdir}/Ancient_DNA/samtools/faidx" }, + enabled: false + ] + } +} \ No newline at end of file diff --git a/modules.json b/modules.json index 6dd73824..854ee864 100644 --- a/modules.json +++ b/modules.json @@ -4,13 +4,13 @@ "repos": { "nf-core/modules": { "bcftools/consensus": { - "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "bcftools/index": { - "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "bcftools/view": { - "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "fastp": { "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" @@ -19,22 +19,22 @@ "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, "freebayes": { - "git_sha": "9767b081b9d808d7b6671d34d48c70afb436a5f1" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" }, - "pydamage/analyze": { + "prokka": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" }, + "pydamage/analyze": { + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + }, "pydamage/filter": { - "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "samtools/faidx": { - "git_sha": "3aacd46da2b221ed47aaa05c413a828538d2c2ae" - }, - "prokka": { - "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" } } } diff --git a/modules/nf-core/modules/bcftools/consensus/functions.nf b/modules/nf-core/modules/bcftools/consensus/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/bcftools/consensus/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/bcftools/consensus/main.nf b/modules/nf-core/modules/bcftools/consensus/main.nf index 954b0eb8..9b9384a6 100644 --- a/modules/nf-core/modules/bcftools/consensus/main.nf +++ b/modules/nf-core/modules/bcftools/consensus/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process BCFTOOLS_CONSENSUS { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' - } else { - container 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : + 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" input: tuple val(meta), path(vcf), path(tbi), path(fasta) @@ -26,15 +15,16 @@ process BCFTOOLS_CONSENSUS { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" """ - cat $fasta | bcftools consensus $vcf $options.args > ${prefix}.fa + cat $fasta | bcftools consensus $vcf $args > ${prefix}.fa header=\$(head -n 1 ${prefix}.fa | sed 's/>//g') sed -i 's/\${header}/${meta.id}/g' ${prefix}.fa cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + "${task.process}": + bcftools: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') END_VERSIONS """ } diff --git a/modules/nf-core/modules/bcftools/index/functions.nf b/modules/nf-core/modules/bcftools/index/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/bcftools/index/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/bcftools/index/main.nf b/modules/nf-core/modules/bcftools/index/main.nf index d67614d8..8f40c683 100644 --- a/modules/nf-core/modules/bcftools/index/main.nf +++ b/modules/nf-core/modules/bcftools/index/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process BCFTOOLS_INDEX { tag "$meta.id" label 'process_low' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0" - } else { - container "quay.io/biocontainers/bcftools:1.13--h3a49de5_0" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : + 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" input: tuple val(meta), path(vcf) @@ -27,18 +16,19 @@ process BCFTOOLS_INDEX { path "versions.yml" , emit: version script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" """ bcftools \\ index \\ - $options.args \\ + $args \\ --threads $task.cpus \\ $vcf cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + "${task.process}": + bcftools: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') END_VERSIONS """ } diff --git a/modules/nf-core/modules/bcftools/view/functions.nf b/modules/nf-core/modules/bcftools/view/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/bcftools/view/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/bcftools/view/main.nf b/modules/nf-core/modules/bcftools/view/main.nf index ef72f081..b2cbb580 100644 --- a/modules/nf-core/modules/bcftools/view/main.nf +++ b/modules/nf-core/modules/bcftools/view/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process BCFTOOLS_VIEW { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::bcftools=1.13" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0" - } else { - container "quay.io/biocontainers/bcftools:1.13--h3a49de5_0" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : + 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" input: tuple val(meta), path(vcf), path(index) @@ -29,25 +18,24 @@ process BCFTOOLS_VIEW { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" def regions_file = regions ? "--regions-file ${regions}" : "" def targets_file = targets ? "--targets-file ${targets}" : "" def samples_file = samples ? "--samples-file ${samples}" : "" - - """ bcftools view \\ --output ${prefix}.vcf.gz \\ ${regions_file} \\ ${targets_file} \\ ${samples_file} \\ - $options.args \\ + $args \\ --threads $task.cpus \\ ${vcf} cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') + "${task.process}": + bcftools: \$(bcftools --version 2>&1 | head -n1 | sed 's/^.*bcftools //; s/ .*\$//') END_VERSIONS """ } diff --git a/modules/nf-core/modules/freebayes/functions.nf b/modules/nf-core/modules/freebayes/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/freebayes/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/freebayes/main.nf b/modules/nf-core/modules/freebayes/main.nf index 0b23dc40..b9a63d02 100644 --- a/modules/nf-core/modules/freebayes/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -1,38 +1,28 @@ -// Import generic module functions -include { initOptions; saveFiles; getProcessName; getSoftwareName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process FREEBAYES { tag "$meta.id" label 'process_low' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::freebayes=1.3.5" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/freebayes:1.3.5--py38ha193a2f_3" - } else { - container "quay.io/biocontainers/freebayes:1.3.5--py38ha193a2f_3" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/freebayes:1.3.5--py38ha193a2f_3' : + 'quay.io/biocontainers/freebayes:1.3.5--py38ha193a2f_3' }" input: tuple val(meta), path(input_1), path(input_1_index), path(input_2), path(input_2_index) path fasta - path fai + path fasta_fai path targets path samples path populations path cnv output: - tuple val(meta), path("*.vcf.gz") , emit: vcf - path "versions.yml" , emit: versions + tuple val(meta), path("*.vcf.gz"), emit: vcf + path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" def input = input_2 ? "${input_1} ${input_2}" : "${input_1}" def targets_file = targets ? "--target ${targets}" : "" def samples_file = samples ? "--samples ${samples}" : "" @@ -42,20 +32,20 @@ process FREEBAYES { if (task.cpus > 1) { """ freebayes-parallel \\ - <(fasta_generate_regions.py ${fasta}.fai 10000) ${task.cpus} \\ + <(fasta_generate_regions.py $fasta_fai 10000) $task.cpus \\ -f $fasta \\ $targets_file \\ $samples_file \\ $populations_file \\ $cnv_file \\ - $options.args \\ + $args \\ $input > ${prefix}.vcf gzip --no-name ${prefix}.vcf cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) + "${task.process}": + freebayes: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) END_VERSIONS """ @@ -67,14 +57,14 @@ process FREEBAYES { $samples_file \\ $populations_file \\ $cnv_file \\ - $options.args \\ + $args \\ $input > ${prefix}.vcf gzip --no-name ${prefix}.vcf cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) + "${task.process}": + freebayes: \$(echo \$(freebayes --version 2>&1) | sed 's/version:\s*v//g' ) END_VERSIONS """ } diff --git a/modules/nf-core/modules/freebayes/meta.yml b/modules/nf-core/modules/freebayes/meta.yml index 75d44826..abba1daa 100644 --- a/modules/nf-core/modules/freebayes/meta.yml +++ b/modules/nf-core/modules/freebayes/meta.yml @@ -36,10 +36,10 @@ input: type: file description: reference fasta file pattern: ".{fa,fa.gz,fasta,fasta.gz}" - - fai: + - fasta_fai: type: file description: reference fasta file index - pattern: "*.fai" + pattern: "*.{fa,fasta}.fai" - targets: type: file description: Optional - Limit analysis to targets listed in this BED-format FILE. @@ -55,8 +55,7 @@ input: - cnv: type: file description: | - A copy number map BED file, which has - either a sample-level ploidy: + A copy number map BED file, which has either a sample-level ploidy: sample_name copy_number or a region-specific format: seq_name start end sample_name copy_number diff --git a/modules/nf-core/modules/pydamage/analyze/functions.nf b/modules/nf-core/modules/pydamage/analyze/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/pydamage/analyze/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/pydamage/analyze/main.nf b/modules/nf-core/modules/pydamage/analyze/main.nf index 9cfb8a1a..c55616db 100644 --- a/modules/nf-core/modules/pydamage/analyze/main.nf +++ b/modules/nf-core/modules/pydamage/analyze/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process PYDAMAGE_ANALYZE { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0" - } else { - container "quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0' : + 'quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0' }" input: tuple val(meta), path(bam), path(bai) @@ -26,17 +15,18 @@ process PYDAMAGE_ANALYZE { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" """ pydamage \\ analyze \\ - $options.args \\ + $args \\ -p $task.cpus \\ $bam cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') + "${task.process}": + pydamage: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') END_VERSIONS """ } diff --git a/modules/nf-core/modules/pydamage/filter/functions.nf b/modules/nf-core/modules/pydamage/filter/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/pydamage/filter/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/pydamage/filter/main.nf b/modules/nf-core/modules/pydamage/filter/main.nf index 6cd7ae7a..2e0afac9 100644 --- a/modules/nf-core/modules/pydamage/filter/main.nf +++ b/modules/nf-core/modules/pydamage/filter/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process PYDAMAGE_FILTER { tag "$meta.id" label 'process_low' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0" - } else { - container "quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0' : + 'quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0' }" input: tuple val(meta), path(csv) @@ -26,17 +15,18 @@ process PYDAMAGE_FILTER { path "versions.yml" , emit: versions script: - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def args = task.ext.args ?: '' + def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" """ pydamage \\ filter \\ - $options.args \\ + $args \\ $csv cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') + "${task.process}": + pydamage: \$(echo \$(pydamage --version 2>&1) | sed -e 's/pydamage, version //g') END_VERSIONS """ } diff --git a/modules/nf-core/modules/samtools/faidx/functions.nf b/modules/nf-core/modules/samtools/faidx/functions.nf deleted file mode 100644 index 85628ee0..00000000 --- a/modules/nf-core/modules/samtools/faidx/functions.nf +++ /dev/null @@ -1,78 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Extract name of module from process name using $task.process -// -def getProcessName(task_process) { - return task_process.tokenize(':')[-1] -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - - // Do not publish versions.yml unless running from pytest workflow - if (args.filename.equals('versions.yml') && !System.getenv("NF_CORE_MODULES_TEST")) { - return null - } - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } -} diff --git a/modules/nf-core/modules/samtools/faidx/main.nf b/modules/nf-core/modules/samtools/faidx/main.nf index 80cedeab..c53373a9 100644 --- a/modules/nf-core/modules/samtools/faidx/main.nf +++ b/modules/nf-core/modules/samtools/faidx/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName; getProcessName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process SAMTOOLS_FAIDX { tag "$fasta" label 'process_low' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:[:], publish_by_meta:[]) } - conda (params.enable_conda ? 'bioconda::samtools=1.13' : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/samtools:1.13--h8c37831_0" - } else { - container "quay.io/biocontainers/samtools:1.13--h8c37831_0" - } + conda (params.enable_conda ? "bioconda::samtools=1.14" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.14--hb421002_0' : + 'quay.io/biocontainers/samtools:1.14--hb421002_0' }" input: path fasta @@ -26,11 +15,12 @@ process SAMTOOLS_FAIDX { path "versions.yml", emit: versions script: + def args = task.ext.args ?: '' """ samtools faidx $fasta cat <<-END_VERSIONS > versions.yml - ${getProcessName(task.process)}: - ${getSoftwareName(task.process)}: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') END_VERSIONS """ } From ee20e6ba70c4f5d0e4ac7008e32e71f17aa62d2b Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 3 Dec 2021 15:55:05 +0100 Subject: [PATCH 022/246] capitalize module names --- conf/modules.config | 16 ++++++++-------- modules/nf-core/modules/freebayes/main.nf | 4 ++-- 2 files changed, 10 insertions(+), 10 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 393b0ae1..1617cc0b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -184,7 +184,7 @@ process { saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] - withName: freebayes { + withName: FREEBAYES { ext.args = "-p ${params.freebayes_ploidy}" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/freebayes" }, @@ -192,7 +192,7 @@ process { ] } - withName: bcftools_view { + withName: BCFTOOLS_VIEW { ext.args = "-i '%QUAL>=${params.bcftools_view_variant_quality}'" ext.suffix = ".filtered" publishDir = [ @@ -201,35 +201,35 @@ process { ] } - withName: bcftools_consensus { + withName: BCFTOOLS_CONSENSUS { publishDir = [ path: {"${params.outdir}/Ancient_DNA/variant_calling/consensus" }, pattern: "*.fa" ] } - withName: bcftools_index { + withName: BCFTOOLS_INDEX { ext.args = "-t" publishDir = [ path: {"${params.outdir}/Ancient_DNA/variant_calling/index" }, enabled: false ] } - - withName: pydamage_analyze { + + withName: PYDAMAGE_ANALYZE { publishDir = [ path: {"${params.outdir}/Ancient_DNA/pydamage/analyze" } ] } - withName: pydamage_filter { + withName: PYDAMAGE_FILTER { ext.args = "-t ${params.pydamage_accuracy}" publishDir = [ path: {"${params.outdir}/Ancient_DNA/pydamage/filter" } ] } - withName: samtools_faidx { + withName: SAMTOOLS_FAIDX { publishDir = [ path: {"${params.outdir}/Ancient_DNA/samtools/faidx" }, enabled: false diff --git a/modules/nf-core/modules/freebayes/main.nf b/modules/nf-core/modules/freebayes/main.nf index b9a63d02..0568ca88 100644 --- a/modules/nf-core/modules/freebayes/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -41,7 +41,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - gzip --no-name ${prefix}.vcf + bgzip ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": @@ -60,7 +60,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - gzip --no-name ${prefix}.vcf + bgzip ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": From 3ad95034c796ff1bc31d27f2efde089f75c46a56 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 3 Dec 2021 16:00:57 +0100 Subject: [PATCH 023/246] remove trailing whitespaces --- conf/modules.config | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 1617cc0b..1ca28ca4 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -216,20 +216,20 @@ process { ] } - withName: PYDAMAGE_ANALYZE { + withName: PYDAMAGE_ANALYZE { publishDir = [ path: {"${params.outdir}/Ancient_DNA/pydamage/analyze" } ] } - withName: PYDAMAGE_FILTER { - ext.args = "-t ${params.pydamage_accuracy}" + withName: PYDAMAGE_FILTER { + ext.args = "-t ${params.pydamage_accuracy}" publishDir = [ path: {"${params.outdir}/Ancient_DNA/pydamage/filter" } ] } - withName: SAMTOOLS_FAIDX { + withName: SAMTOOLS_FAIDX { publishDir = [ path: {"${params.outdir}/Ancient_DNA/samtools/faidx" }, enabled: false From 8fa32900567327e086c3c757791e4fc1cbb7f028 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 3 Dec 2021 16:04:32 +0100 Subject: [PATCH 024/246] add final newline --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 1ca28ca4..ee293358 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -235,4 +235,4 @@ process { enabled: false ] } -} \ No newline at end of file +} From a73596b2afa9c43a67d14ce490015b5767cb6d67 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Dec 2021 21:22:14 +0100 Subject: [PATCH 025/246] fix: possible fix to metabat2? --- conf/modules.config | 6 ++-- modules/local/mag_depths.nf | 6 ++-- modules/nf-core/modules/maxbin2/main.nf | 2 +- .../jgisummarizebamcontigdepths/main.nf | 2 +- .../nf-core/modules/metabat2/metabat2/main.nf | 24 ++++++++----- nf-core/modules/gunzip/main.nf | 32 ----------------- nf-core/modules/gunzip/meta.yml | 34 ------------------ subworkflows/local/metabat2_binning.nf | 36 +++++++++---------- 8 files changed, 41 insertions(+), 101 deletions(-) delete mode 100644 nf-core/modules/gunzip/main.nf delete mode 100644 nf-core/modules/gunzip/meta.yml diff --git a/conf/modules.config b/conf/modules.config index c75819d9..37428381 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -182,16 +182,16 @@ process { withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { publishDir = [ - path: { "${params.outdir}/GenomeBinning/" }, + path: { "${params.outdir}/GenomeBinning/METABAT2" }, mode: 'copy', pattern: '*_depth.txt.gz' ] - ext.suffix = { "_${meta.assembler}_depth" } + ext.suffix = { "${meta.assembler}-${meta.id}-depth" } } withName: METABAT2_METABAT2 { publishDir = [ - path: { "${params.outdir}/GenomeBinning/" }, + path: { "${params.outdir}/GenomeBinning/METABAT2" }, mode: 'copy' ] ext.args = [ diff --git a/modules/local/mag_depths.nf b/modules/local/mag_depths.nf index 533496b5..c41bc3e8 100644 --- a/modules/local/mag_depths.nf +++ b/modules/local/mag_depths.nf @@ -28,9 +28,9 @@ process MAG_DEPTHS { script: def software = getSoftwareName(task.process) """ - get_mag_depths.py --bins *.fa.gz \ - --depths ${contig_depths} \ - --assembly_name "${meta.assembler}-${meta.id}" \ + get_mag_depths.py --bins ${bins} \\ + --depths ${contig_depths} \\ + --assembly_name "${meta.assembler}-${meta.id}" \\ --out "${meta.assembler}-${meta.id}-binDepths.tsv" """ } diff --git a/modules/nf-core/modules/maxbin2/main.nf b/modules/nf-core/modules/maxbin2/main.nf index e13af704..4d384391 100644 --- a/modules/nf-core/modules/maxbin2/main.nf +++ b/modules/nf-core/modules/maxbin2/main.nf @@ -23,7 +23,7 @@ process MAXBIN2 { script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" def associate_files = reads ? "-reads $reads" : "-abund $abund" """ run_MaxBin.pl \\ diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf index e35d6715..4a5869b6 100644 --- a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf @@ -16,7 +16,7 @@ process METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" """ export OMP_NUM_THREADS=$task.cpus diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index 1b180fc8..6c0781db 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -11,12 +11,12 @@ process METABAT2_METABAT2 { tuple val(meta), path(fasta), path(depth) output: - tuple val(meta), path("bins/*.fa.gz") , optional:true , emit: fasta - tuple val(meta), path("*.tooShort.fa.gz") , optional:true , emit: tooshort - tuple val(meta), path("*.lowDepth.fa.gz") , optional:true , emit: lowdepth - tuple val(meta), path("*.unbinned.fa.gz") , optional:true , emit: unbinned - tuple val(meta), path("*.tsv.gz") , optional:true , emit: membership - path "versions.yml" , emit: versions + tuple val(meta), path("*.tooShort.fa.gz") , optional:true , emit: tooshort + tuple val(meta), path("*.lowDepth.fa.gz") , optional:true , emit: lowdepth + tuple val(meta), path("*.unbinned.fa.gz") , optional:true , emit: unbinned + tuple val(meta), path("*.tsv.gz") , optional:true , emit: membership + tuple val(meta), path("bins/*.fa.gz") , optional:true , emit: fasta + path "versions.yml" , emit: versions script: def args = task.ext.args ?: '' @@ -37,9 +37,17 @@ process METABAT2_METABAT2 { mv metabat2/${prefix} ${prefix}.tsv mv metabat2 bins - gzip ${prefix}.tsv + gzip ${prefix}.tsv gzip bins/*.fa - mv bins/*.{tooShort,lowDepth,unbinned}.fa.gz . + + ## Some bash trickeryy to successfully execute `mv`, even if no contigs + ## discarded. Thanks to Stéphane Chazelas https://unix.stackexchange.com/a/522283/478356 + ( + shopt -s nullglob + shopt -u failglob + files=(bins/*{lowDepth,tooShort,unbinned}.fa.gz) + ((\${#files[@]} == 0)) || mv -- "\${files[@]}" . + ) cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/nf-core/modules/gunzip/main.nf b/nf-core/modules/gunzip/main.nf deleted file mode 100644 index ba9aae48..00000000 --- a/nf-core/modules/gunzip/main.nf +++ /dev/null @@ -1,32 +0,0 @@ -process GUNZIP { - tag "$archive" - label 'process_low' - - conda (params.enable_conda ? "conda-forge::sed=4.7" : null) - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" - - input: - tuple val(meta), path(archive) - - output: - tuple val(meta), path("$gunzip"), emit: gunzip - path "versions.yml" , emit: versions - - script: - def args = task.ext.args ?: '' - gunzip = archive.collect { it.toString() - '.gz' } - """ - echo "${archive.toString() - '.gz'}" - gunzip \\ - -f \\ - $args \\ - $archive - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - gunzip: \$(echo \$(gunzip --version 2>&1) | sed 's/^.*(gzip) //; s/ Copyright.*\$//') - END_VERSIONS - """ -} diff --git a/nf-core/modules/gunzip/meta.yml b/nf-core/modules/gunzip/meta.yml deleted file mode 100644 index ea1f1546..00000000 --- a/nf-core/modules/gunzip/meta.yml +++ /dev/null @@ -1,34 +0,0 @@ -name: gunzip -description: Compresses and decompresses files. -keywords: - - gunzip - - compression -tools: - - gunzip: - description: | - gzip is a file format and a software application used for file compression and decompression. - documentation: https://www.gnu.org/software/gzip/manual/gzip.html - licence: ['GPL-3.0-or-later'] -input: - - meta: - type: map - description: | - Optional groovy Map containing meta information - e.g. [ id:'test', single_end:false ] - - archive: - type: file - description: File to be compressed/uncompressed - pattern: "*.*" -output: - - gunzip: - type: file - description: Compressed/uncompressed file - pattern: "*.*" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" -authors: - - "@joseespinosa" - - "@drpatelh" - - "@jfy133" diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 41feb846..7e513c07 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -33,9 +33,7 @@ workflow METABAT2_BINNING { ch_summarizedepth_input = assemblies .map { meta, assembly, bams, bais -> [ meta, bams, bais ] } - METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( - ch_summarizedepth_input - ) + METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) ch_metabat2_input = assemblies .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) @@ -43,24 +41,27 @@ workflow METABAT2_BINNING { METABAT2_METABAT2 ( ch_metabat2_input ) + METABAT2_METABAT2.out.fasta.dump(tag:"fasta_channel_metabat2_output") + // split FASTQ - SPLIT_FASTQ ( - METABAT2_METABAT2.out.unbinned - ) + SPLIT_FASTQ ( METABAT2_METABAT2.out.unbinned ) + + // decompress main bins for downstream, have to separate and re-group due to limitation of GUNZIP + METABAT2_METABAT2.out.fasta + .transpose() + .set { ch_metabat2_results_transposed } - // decompress main bins for downstream - GUNZIP ( - METABAT2_METABAT2.out.fasta - ) + GUNZIP ( ch_metabat2_results_transposed ) + + GUNZIP.out.gunzip.groupTuple(by: 0).set{ ch_metabat_results_gunzipped } // Compute bin depths for different samples (according to `binning_map_mode`) - ch_depth_input = METABAT2_METABAT2.out.fasta + ch_depth_input = ch_metabat_results_gunzipped .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) .dump(tag:"pre_for_mag_depths") - MAG_DEPTHS ( - ch_depth_input - ) + MAG_DEPTHS ( ch_depth_input ) + // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples ch_sample_groups = reads @@ -70,15 +71,12 @@ workflow METABAT2_BINNING { ch_mag_depths_plot = MAG_DEPTHS.out.depths .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } - MAG_DEPTHS_PLOT ( - ch_mag_depths_plot, - ch_sample_groups.collect() - ) + MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) emit: - bins = GUNZIP.out.gunzip // TODO this would include discarded FASTAS! need to separate out somehow! Probably in metabat2 module + bins = ch_metabat_results_gunzipped // TODO this would include discarded FASTAS! need to separate out somehow! Probably in metabat2 module unbinned = SPLIT_FASTQ.out.unbinned tooshort = METABAT2_METABAT2.out.tooshort lowdepth = METABAT2_METABAT2.out.lowdepth From 93ca9ae3b560e98d4a53756f828911500c37362c Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 6 Dec 2021 17:27:13 +0100 Subject: [PATCH 026/246] aDNA subworkflow --- conf/modules.config | 2 +- conf/test_ancient_dna.config | 2 +- modules.json | 2 +- .../nf-core/modules/samtools/faidx/main.nf | 6 +- .../nf-core/modules/samtools/faidx/meta.yml | 10 +++ subworkflows/local/ancient_dna.nf | 13 +++- workflows/mag.nf | 73 ++++++++++++------- 7 files changed, 74 insertions(+), 34 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 61440e1a..0d48ab04 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -97,7 +97,7 @@ params { publish_dir = "Assembly" } 'bowtie2_assembly_align' { - args = params.bowtie2_mode + args = params.bowtie2_mode ? params.bowtie2_mode : "" publish_files = ['log':''] publish_by_meta = ['assembler', 'QC', 'id'] publish_dir = "Assembly" diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config index 798ad500..1f7081ba 100644 --- a/conf/test_ancient_dna.config +++ b/conf/test_ancient_dna.config @@ -30,7 +30,7 @@ params { gtdb = false ancient_dna = true binning_map_mode = 'own' - skip_spades = true + skip_spades = false skip_spadeshybrid = true bcftools_view_variant_quality = 0 bowtie2_mode = "--very-sensitive -N 1" diff --git a/modules.json b/modules.json index 00350981..cfbcbf59 100644 --- a/modules.json +++ b/modules.json @@ -46,7 +46,7 @@ "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "samtools/faidx": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e0aa89141ffecb5f54d230f7ea46de242b74e084" } } } diff --git a/modules/nf-core/modules/samtools/faidx/main.nf b/modules/nf-core/modules/samtools/faidx/main.nf index c53373a9..d8308b03 100644 --- a/modules/nf-core/modules/samtools/faidx/main.nf +++ b/modules/nf-core/modules/samtools/faidx/main.nf @@ -8,11 +8,11 @@ process SAMTOOLS_FAIDX { 'quay.io/biocontainers/samtools:1.14--hb421002_0' }" input: - path fasta + tuple val(meta), path(fasta) output: - path "*.fai" , emit: fai - path "versions.yml", emit: versions + tuple val(meta), path ("*.fai") , emit: fai + path "versions.yml" , emit: versions script: def args = task.ext.args ?: '' diff --git a/modules/nf-core/modules/samtools/faidx/meta.yml b/modules/nf-core/modules/samtools/faidx/meta.yml index 16c0b334..bae97a39 100644 --- a/modules/nf-core/modules/samtools/faidx/meta.yml +++ b/modules/nf-core/modules/samtools/faidx/meta.yml @@ -14,11 +14,21 @@ tools: doi: 10.1093/bioinformatics/btp352 licence: ['MIT'] input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] - fasta: type: file description: FASTA file pattern: "*.{fa,fasta}" output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] - fai: type: file description: FASTA index file diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index 73ad5910..c770653f 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -20,8 +20,17 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { main: PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) - FAIDX(input.map { item -> [ item[1] ] }) - FREEBAYES (input.map { item -> [item[0], item[2], item[3], [], []] }, input.map { item -> [ item[1] ] }, FAIDX.out.fai, [], [], [], [] ) + FAIDX(input.map { item -> [ item[0], item[1] ] }) + input.join(FAIDX.out.fai) + .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] + FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], []] }, + freebayes_input.map { item -> item[1] }, + freebayes_input.map { item -> item[4] }, + [], + [], + [], + [] ) + BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) BCFTOOLS_INDEX_POST(BCFTOOLS_VIEW.out.vcf) diff --git a/workflows/mag.nf b/workflows/mag.nf index b6c29fca..a16e01b3 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -91,11 +91,12 @@ include { MULTIQC } from '../modules // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' -include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' //addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) -include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) -include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) +include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' //addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) +include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) +include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' /* ======================================================================================== @@ -487,25 +488,56 @@ workflow MAG { /* ================================================================================ - Binning + Binning preparation ================================================================================ */ + ch_bowtie2_assembly_multiqc = Channel.empty() ch_busco_summary = Channel.empty() ch_busco_multiqc = Channel.empty() - if (!params.skip_binning){ + - BINNING_PREPARATION ( - ch_assemblies, - ch_short_reads - ) + BINNING_PREPARATION ( + ch_assemblies, + ch_short_reads + ) - METABAT2_BINNING ( - BINNING_PREPARATION.out.grouped_mappings, - ch_short_reads - ) + /* + ================================================================================ + Ancient DNA + ================================================================================ + */ + + if (params.ancient_dna){ + ANCIENT_DNA_ASSEMLY_VALIDATION(BINNING_PREPARATION.out.grouped_mappings) + ch_software_versions = ch_software_versions.mix(ANCIENT_DNA_ASSEMLY_VALIDATION.out.versions.ifEmpty(null)) + } + + /* + ================================================================================ + Binning + ================================================================================ + */ + + if (!params.skip_binning){ + + if (params.ancient_dna) { + METABAT2_BINNING ( + BINNING_PREPARATION.out.grouped_mappings + .join(ANCIENT_DNA_ASSEMLY_VALIDATION.out.contigs_recalled) + .map{ it -> [ it[0], it[4], it[2], it[3] ] }, // [meta, contigs_recalled, bam, bais] + ch_short_reads + ) + } else { + METABAT2_BINNING ( + BINNING_PREPARATION.out.grouped_mappings, + ch_short_reads + ) + } + + ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_software_versions = ch_software_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first().ifEmpty(null)) ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.metabat2_version.first().ifEmpty(null)) @@ -599,17 +631,6 @@ workflow MAG { } } - /* - ================================================================================ - Ancient DNA - ================================================================================ - */ - - if (params.ancient_dna){ - ANCIENT_DNA_ASSEMLY_VALIDATION(METABAT2_BINNING.out.assembly_mappings) - ch_software_versions = ch_software_versions.mix(ANCIENT_DNA_ASSEMLY_VALIDATION.out.versions.ifEmpty(null)) - } - // // MODULE: Pipeline reporting // From 2cd6af01ea71ce161049634853cdab225aad493d Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 7 Dec 2021 09:46:31 +0100 Subject: [PATCH 027/246] fix: temp. metabat2 fix (will reinstall when merged) --- modules/nf-core/modules/metabat2/metabat2/main.nf | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index 6c0781db..e83a3069 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -40,8 +40,9 @@ process METABAT2_METABAT2 { gzip ${prefix}.tsv gzip bins/*.fa - ## Some bash trickeryy to successfully execute `mv`, even if no contigs - ## discarded. Thanks to Stéphane Chazelas https://unix.stackexchange.com/a/522283/478356 + ## Some bash trickery to successfully execute `mv`, even when no contigs are + ## actually discarded. + ## Thanks to Stéphane Chazelas https://unix.stackexchange.com/a/522283/478356 ( shopt -s nullglob shopt -u failglob From ba783c87250cb07e9d8fbad95ad61f492bace71e Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 7 Dec 2021 09:47:00 +0100 Subject: [PATCH 028/246] fix: fix metabat2 and update split_fastq to match original structure --- conf/base.config | 2 +- conf/modules.config | 9 ++++++++- docs/output.md | 4 ++-- modules/local/split_fastq.nf | 8 ++++---- modules/nf-core/modules/metabat2/metabat2/main.nf | 15 +++------------ 5 files changed, 18 insertions(+), 20 deletions(-) diff --git a/conf/base.config b/conf/base.config index cd177b76..98b69c1d 100644 --- a/conf/base.config +++ b/conf/base.config @@ -143,7 +143,7 @@ process { time = { check_max (8.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in [143,137,104,134,139,247] ? 'retry' : 'finish' } } - withName: METABAT2 { + withName: METABAT2_METABAT2 { cpus = { check_max (8 * task.attempt, 'cpus' ) } memory = { check_max (20.GB * task.attempt, 'memory' ) } time = { check_max (8.h * task.attempt, 'time' ) } diff --git a/conf/modules.config b/conf/modules.config index 37428381..0ea14576 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -191,7 +191,7 @@ process { withName: METABAT2_METABAT2 { publishDir = [ - path: { "${params.outdir}/GenomeBinning/METABAT2" }, + path: { "${params.outdir}/GenomeBinning/METABAT2/" }, mode: 'copy' ] ext.args = [ @@ -201,4 +201,11 @@ process { ].join(' ').trim() } + withName: SPLIT_FASTQ { + publishedDir = [ + path: "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" } + mode: 'copy' + ] + } + } diff --git a/docs/output.md b/docs/output.md index 3e77ca31..6a6275e7 100644 --- a/docs/output.md +++ b/docs/output.md @@ -253,8 +253,8 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig Output files * `GenomeBinning/MetaBAT2/` - * `[assembler]-[sample/group].*.fa`: Genome bins retrieved from input assembly - * `[assembler]-[sample/group].unbinned.*.fa`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported + * `bins/[assembler]-[sample/group].*.fa`: Genome bins retrieved from input assembly + * `discarded/[assembler]-[sample/group].{unbinned,lowDepth,tooShort}.*.fa`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported diff --git a/modules/local/split_fastq.nf b/modules/local/split_fastq.nf index 2d1d047f..5cc19cb4 100644 --- a/modules/local/split_fastq.nf +++ b/modules/local/split_fastq.nf @@ -14,15 +14,15 @@ process SPLIT_FASTQ { tuple val(meta), path(unbinned) output: - path "${meta.assembler}/unbinned/" , emit: unbinned + path "${meta.assembler}/discarded/" , emit: unbinned script: """ # save unbinned contigs above thresholds into individual files, dump others in one file split_fasta.py $unbinned ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} - mkdir -p ${meta.assembler}/unbinned/ - mv *.fa ${meta.assembler}/unbinned/ - gzip ${meta.assembler}/unbinned/* + mkdir -p ${meta.assembler}/discarded/ + mv *.fa ${meta.assembler}/discarded/ + gzip ${meta.assembler}/discarded/* """ } diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index e83a3069..a8af0ae9 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -20,7 +20,7 @@ process METABAT2_METABAT2 { script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" def decompress_depth = depth ? "gzip -d -f $depth" : "" def depth_file = depth ? "-a ${depth.baseName}" : "" """ @@ -38,17 +38,8 @@ process METABAT2_METABAT2 { mv metabat2 bins gzip ${prefix}.tsv - gzip bins/*.fa - - ## Some bash trickery to successfully execute `mv`, even when no contigs are - ## actually discarded. - ## Thanks to Stéphane Chazelas https://unix.stackexchange.com/a/522283/478356 - ( - shopt -s nullglob - shopt -u failglob - files=(bins/*{lowDepth,tooShort,unbinned}.fa.gz) - ((\${#files[@]} == 0)) || mv -- "\${files[@]}" . - ) + find ./bins/ -name "*.fa" -type f | xargs -t -n 1 bgzip -@ ${task.cpus} + find ./bins/ -name "*[lowDepth,tooShort,unbinned].fa.gz" -type f -exec mv {} . \\; cat <<-END_VERSIONS > versions.yml "${task.process}": From 9677a61358f5082ff93d4da384a0bf7b8806378e Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 7 Dec 2021 09:49:28 +0100 Subject: [PATCH 029/246] fix: missing bracket --- conf/modules.config | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 0ea14576..448c5527 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -191,7 +191,7 @@ process { withName: METABAT2_METABAT2 { publishDir = [ - path: { "${params.outdir}/GenomeBinning/METABAT2/" }, + path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, mode: 'copy' ] ext.args = [ @@ -202,8 +202,8 @@ process { } withName: SPLIT_FASTQ { - publishedDir = [ - path: "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" } + publishDir = [ + path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded" }, mode: 'copy' ] } From a8478e7017261af2b10d8a314bf41812eee046b1 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 13 Dec 2021 16:12:04 +0100 Subject: [PATCH 030/246] fix: update metabat2, restructure bin dirs, still stuck on depth publishing --- bin/split_fasta.py | 6 ++- conf/modules.config | 38 +++++++++++++------ modules.json | 2 +- modules/local/split_fastq.nf | 8 ++-- .../modules/metabat2/metabat2/meta.yml | 13 +++++++ subworkflows/local/metabat2_binning.nf | 30 ++++++++++++--- 6 files changed, 73 insertions(+), 24 deletions(-) diff --git a/bin/split_fasta.py b/bin/split_fasta.py index c7c6e6bc..c9eb6886 100755 --- a/bin/split_fasta.py +++ b/bin/split_fasta.py @@ -18,7 +18,11 @@ min_length_to_retain_contig = int(argv[4]) # Base name for file output -out_base = (os.path.splitext(input_file)[0]) +if input_file.endswith('.gz'): + rm_ext=os.path.splitext(input_file)[0] + out_base = (os.path.splitext(rm_ext)[0]) +else: + out_base = (os.path.splitext(input_file)[0]) # Data structures to separate and store sequences df_above_threshold = pd.DataFrame(columns=['id','seq','length']) diff --git a/conf/modules.config b/conf/modules.config index 448c5527..5bd7269b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -105,10 +105,10 @@ params { publish_files = false } 'mag_depths_plot' { - publish_dir = "GenomeBinning" + publish_dir = "GenomeBinning/depths" } 'mag_depths_summary' { - publish_dir = "GenomeBinning" + publish_dir = "GenomeBinning/depths" } 'busco_db_preparation' { publish_files = ['tar.gz':''] @@ -174,15 +174,15 @@ params { // nf-core modules DSL2 syntax v2.0 process { - publishDir = [ - path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] + // publishDir = [ + // path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + // mode: params.publish_dir_mode, + // saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + //] withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { publishDir = [ - path: { "${params.outdir}/GenomeBinning/METABAT2" }, + path: { "${params.outdir}/GenomeBinning/MetaBAT2" }, mode: 'copy', pattern: '*_depth.txt.gz' ] @@ -191,9 +191,23 @@ process { withName: METABAT2_METABAT2 { publishDir = [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, - mode: 'copy' + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, + mode: 'copy', + pattern: 'bins/*.fa.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded_contigs" }, + mode: 'copy', + pattern: '*tooShort.fa.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded_contigs" }, + mode: 'copy', + pattern: '*lowDepth.fa.gz' + ] ] + ext.prefix = { "${meta.assembler}-${meta.id}" } ext.args = [ "-m ${params.min_contig_size}", "--unbinned", @@ -203,8 +217,8 @@ process { withName: SPLIT_FASTQ { publishDir = [ - path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded" }, - mode: 'copy' + path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, + mode: 'copy', ] } diff --git a/modules.json b/modules.json index dad8c6eb..8357e4bb 100644 --- a/modules.json +++ b/modules.json @@ -19,7 +19,7 @@ "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "metabat2/metabat2": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "be8528998134132c6f4823f627bbb0aec6a51747" }, "prodigal": { "git_sha": "49da8642876ae4d91128168cd0db4f1c858d7792" diff --git a/modules/local/split_fastq.nf b/modules/local/split_fastq.nf index 5cc19cb4..20c314a1 100644 --- a/modules/local/split_fastq.nf +++ b/modules/local/split_fastq.nf @@ -14,15 +14,15 @@ process SPLIT_FASTQ { tuple val(meta), path(unbinned) output: - path "${meta.assembler}/discarded/" , emit: unbinned + path "unbinned/" , emit: unbinned script: """ # save unbinned contigs above thresholds into individual files, dump others in one file split_fasta.py $unbinned ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} - mkdir -p ${meta.assembler}/discarded/ - mv *.fa ${meta.assembler}/discarded/ - gzip ${meta.assembler}/discarded/* + mkdir -p unbinned/ + mv *.fa unbinned/ + gzip unbinned/* """ } diff --git a/modules/nf-core/modules/metabat2/metabat2/meta.yml b/modules/nf-core/modules/metabat2/metabat2/meta.yml index a7f3a7ff..0ec07b02 100644 --- a/modules/nf-core/modules/metabat2/metabat2/meta.yml +++ b/modules/nf-core/modules/metabat2/metabat2/meta.yml @@ -46,6 +46,18 @@ output: type: file description: Bins created from assembled contigs in fasta file pattern: "*.fa.gz" + - tooshort: + type: file + description: Contigs that did not pass length filtering + pattern: "*.tooShort.fa.gz" + - lowdepth: + type: file + description: Contigs that did not have sufficient depth for binning + pattern: "*.lowDepth.fa.gz" + - unbinned: + type: file + description: Contigs that pass length and depth filtering but could not be binned + pattern: "*.unbinned.fa.gz" - membership: type: file description: cluster memberships as a matrix format. @@ -54,3 +66,4 @@ output: authors: - "@maxibor" + - "@jfy133" diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 7e513c07..3de4fe76 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -35,15 +35,33 @@ workflow METABAT2_BINNING { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) + METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth + .dump(tag: "output_jgi") + .map { it -> + it[0]['binner'] = 'METABAT2' + + [ it[0], it[1] ] + } + .dump(tag: "output_jgi_updated") + .set { ch_metabat_depths } + ch_metabat2_input = assemblies - .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) - . map { it -> [ it[0], it[1], it[4] ] } + .dump(tag: "metabat2_raw") + .map { it -> + it[0]['binner'] = 'METABAT2' + + [ it[0], it[1], it[2], it[3] ] + } + .join( ch_metabat_depths, by: 0 ) + .map { it -> + [ it[0], it[1], it[4]] + } + .dump(tag: "metabat2_input_updated") METABAT2_METABAT2 ( ch_metabat2_input ) - METABAT2_METABAT2.out.fasta.dump(tag:"fasta_channel_metabat2_output") - // split FASTQ + METABAT2_METABAT2.out.unbinned.dump(tag: "input_splitfastq") SPLIT_FASTQ ( METABAT2_METABAT2.out.unbinned ) // decompress main bins for downstream, have to separate and re-group due to limitation of GUNZIP @@ -57,8 +75,8 @@ workflow METABAT2_BINNING { // Compute bin depths for different samples (according to `binning_map_mode`) ch_depth_input = ch_metabat_results_gunzipped + .dump(tag: "depth_input") .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) - .dump(tag:"pre_for_mag_depths") MAG_DEPTHS ( ch_depth_input ) @@ -76,7 +94,7 @@ workflow METABAT2_BINNING { MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) emit: - bins = ch_metabat_results_gunzipped // TODO this would include discarded FASTAS! need to separate out somehow! Probably in metabat2 module + bins = ch_metabat_results_gunzipped unbinned = SPLIT_FASTQ.out.unbinned tooshort = METABAT2_METABAT2.out.tooshort lowdepth = METABAT2_METABAT2.out.lowdepth From d5a0eea13ca19bd487645981b7a0800ccd144710 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Tue, 14 Dec 2021 16:42:57 +0000 Subject: [PATCH 031/246] Template update for nf-core/tools version 2.2 --- .gitattributes | 2 + .github/CONTRIBUTING.md | 38 ++------ .github/ISSUE_TEMPLATE/bug_report.md | 63 ------------- .github/ISSUE_TEMPLATE/bug_report.yml | 52 ++++++++++ .github/ISSUE_TEMPLATE/config.yml | 1 - .github/ISSUE_TEMPLATE/feature_request.md | 32 ------- .github/ISSUE_TEMPLATE/feature_request.yml | 11 +++ .github/workflows/awsfulltest.yml | 8 +- .github/workflows/awstest.yml | 10 +- .github/workflows/ci.yml | 23 +++-- .github/workflows/linting_comment.yml | 1 + CHANGELOG.md | 2 +- CITATIONS.md | 2 +- README.md | 13 ++- assets/multiqc_config.yaml | 2 +- assets/nf-core-mag_logo.png | Bin 15549 -> 0 bytes assets/nf-core-mag_logo_light.png | 9 ++ assets/sendmail_template.txt | 4 +- bin/scrape_software_versions.py | 36 ------- conf/base.config | 3 + conf/modules.config | 55 ++++++----- conf/test.config | 4 +- docs/images/nf-core-mag_logo.png | Bin 30598 -> 0 bytes docs/images/nf-core-mag_logo_dark.png | 11 +++ docs/images/nf-core-mag_logo_light.png | Bin 0 -> 64765 bytes docs/output.md | 2 +- docs/usage.md | 36 ------- lib/NfcoreSchema.groovy | 26 +++-- lib/NfcoreTemplate.groovy | 30 ++---- lib/Utils.groovy | 7 -- lib/WorkflowMain.groovy | 6 +- modules.json | 9 +- modules/local/functions.nf | 68 ------------- modules/local/get_software_versions.nf | 33 ------- modules/local/samplesheet_check.nf | 24 ++--- .../custom/dumpsoftwareversions/main.nf | 21 +++++ .../custom/dumpsoftwareversions/meta.yml | 34 +++++++ .../templates/dumpsoftwareversions.py | 89 ++++++++++++++++++ modules/nf-core/modules/fastqc/functions.nf | 68 ------------- modules/nf-core/modules/fastqc/main.nf | 39 ++++---- modules/nf-core/modules/fastqc/meta.yml | 7 +- modules/nf-core/modules/multiqc/functions.nf | 68 ------------- modules/nf-core/modules/multiqc/main.nf | 31 +++--- modules/nf-core/modules/multiqc/meta.yml | 7 +- nextflow.config | 33 +++---- nextflow_schema.json | 29 ------ subworkflows/local/input_check.nf | 8 +- workflows/mag.nf | 44 +++------ 48 files changed, 430 insertions(+), 671 deletions(-) delete mode 100644 .github/ISSUE_TEMPLATE/bug_report.md create mode 100644 .github/ISSUE_TEMPLATE/bug_report.yml delete mode 100644 .github/ISSUE_TEMPLATE/feature_request.md create mode 100644 .github/ISSUE_TEMPLATE/feature_request.yml delete mode 100644 assets/nf-core-mag_logo.png create mode 100644 assets/nf-core-mag_logo_light.png delete mode 100755 bin/scrape_software_versions.py delete mode 100644 docs/images/nf-core-mag_logo.png create mode 100644 docs/images/nf-core-mag_logo_dark.png create mode 100644 docs/images/nf-core-mag_logo_light.png delete mode 100644 modules/local/functions.nf delete mode 100644 modules/local/get_software_versions.nf create mode 100644 modules/nf-core/modules/custom/dumpsoftwareversions/main.nf create mode 100644 modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml create mode 100644 modules/nf-core/modules/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py delete mode 100644 modules/nf-core/modules/fastqc/functions.nf delete mode 100644 modules/nf-core/modules/multiqc/functions.nf diff --git a/.gitattributes b/.gitattributes index 7fe55006..050bb120 100644 --- a/.gitattributes +++ b/.gitattributes @@ -1 +1,3 @@ *.config linguist-language=nextflow +modules/nf-core/** linguist-generated +subworkflows/nf-core/** linguist-generated diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index ad41f31d..be1d64be 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -68,16 +68,13 @@ If you wish to contribute a new step, please use the following coding standards: 1. Define the corresponding input channel into your new process from the expected previous process channel 2. Write the process block (see below). 3. Define the output channel if needed (see below). -4. Add any new flags/options to `nextflow.config` with a default (see below). -5. Add any new flags/options to `nextflow_schema.json` with help text (with `nf-core schema build`). -6. Add any new flags/options to the help message (for integer/text parameters, print to help the corresponding `nextflow.config` parameter). -7. Add sanity checks for all relevant parameters. -8. Add any new software to the `scrape_software_versions.py` script in `bin/` and the version command to the `scrape_software_versions` process in `main.nf`. -9. Do local tests that the new code works properly and as expected. -10. Add a new test command in `.github/workflow/ci.yml`. -11. If applicable add a [MultiQC](https://https://multiqc.info/) module. -12. Update MultiQC config `assets/multiqc_config.yaml` so relevant suffixes, name clean up, General Statistics Table column order, and module figures are in the right order. -13. Optional: Add any descriptions of MultiQC report sections and output files to `docs/output.md`. +4. Add any new parameters to `nextflow.config` with a default (see below). +5. Add any new parameters to `nextflow_schema.json` with help text (via the `nf-core schema build` tool). +6. Add sanity checks and validation for all relevant parameters. +7. Perform local tests to validate that the new code works as expected. +8. If applicable, add a new test command in `.github/workflow/ci.yml`. +9. Update MultiQC config `assets/multiqc_config.yaml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module. +10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`. ### Default values @@ -102,27 +99,6 @@ Please use the following naming schemes, to make it easy to understand what is g If you are using a new feature from core Nextflow, you may bump the minimum required version of nextflow in the pipeline with: `nf-core bump-version --nextflow . [min-nf-version]` -### Software version reporting - -If you add a new tool to the pipeline, please ensure you add the information of the tool to the `get_software_version` process. - -Add to the script block of the process, something like the following: - -```bash - --version &> v_.txt 2>&1 || true -``` - -or - -```bash - --help | head -n 1 &> v_.txt 2>&1 || true -``` - -You then need to edit the script `bin/scrape_software_versions.py` to: - -1. Add a Python regex for your tool's `--version` output (as in stored in the `v_.txt` file), to ensure the version is reported as a `v` and the version number e.g. `v2.1.1` -2. Add a HTML entry to the `OrderedDict` for formatting in MultiQC. - ### Images and figures For overview images and other documents we follow the nf-core [style guidelines and examples](https://nf-co.re/developers/design_guidelines). diff --git a/.github/ISSUE_TEMPLATE/bug_report.md b/.github/ISSUE_TEMPLATE/bug_report.md deleted file mode 100644 index 11b427d2..00000000 --- a/.github/ISSUE_TEMPLATE/bug_report.md +++ /dev/null @@ -1,63 +0,0 @@ ---- -name: Bug report -about: Report something that is broken or incorrect -labels: bug ---- - - - -## Check Documentation - -I have checked the following places for your error: - -- [ ] [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting) -- [ ] [nf-core/mag pipeline documentation](https://nf-co.re/mag/usage) - -## Description of the bug - - - -## Steps to reproduce - -Steps to reproduce the behaviour: - -1. Command line: -2. See error: - -## Expected behaviour - - - -## Log files - -Have you provided the following extra information/files: - -- [ ] The command used to run the pipeline -- [ ] The `.nextflow.log` file - -## System - -- Hardware: -- Executor: -- OS: -- Version - -## Nextflow Installation - -- Version: - -## Container engine - -- Engine: -- version: - -## Additional context - - diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml new file mode 100644 index 00000000..8af04d7f --- /dev/null +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -0,0 +1,52 @@ + +name: Bug report +description: Report something that is broken or incorrect +labels: bug +body: + + - type: markdown + attributes: + value: | + Before you post this issue, please check the documentation: + + - [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting) + - [nf-core/mag pipeline documentation](https://nf-co.re/mag/usage) + + - type: textarea + id: description + attributes: + label: Description of the bug + description: A clear and concise description of what the bug is. + validations: + required: true + + - type: textarea + id: command_used + attributes: + label: Command used and terminal output + description: Steps to reproduce the behaviour. Please paste the command you used to launch the pipeline and the output from your terminal. + render: console + placeholder: | + $ nextflow run ... + + Some output where something broke + + - type: textarea + id: files + attributes: + label: Relevant files + description: | + Please drag and drop the relevant files here. Create a `.zip` archive if the extension is not allowed. + Your verbose log file `.nextflow.log` is often useful _(this is a hidden file in the directory where you launched the pipeline)_ as well as custom Nextflow configuration files. + + - type: textarea + id: system + attributes: + label: System information + description: | + * Nextflow version _(eg. 21.10.3)_ + * Hardware _(eg. HPC, Desktop, Cloud)_ + * Executor _(eg. slurm, local, awsbatch)_ + * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter or Charliecloud)_ + * OS _(eg. CentOS Linux, macOS, Linux Mint)_ + * Version of nf-core/mag _(eg. 1.1, 1.5, 1.8.2)_ diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml index f11d124c..3286c46b 100644 --- a/.github/ISSUE_TEMPLATE/config.yml +++ b/.github/ISSUE_TEMPLATE/config.yml @@ -1,4 +1,3 @@ -blank_issues_enabled: false contact_links: - name: Join nf-core url: https://nf-co.re/join diff --git a/.github/ISSUE_TEMPLATE/feature_request.md b/.github/ISSUE_TEMPLATE/feature_request.md deleted file mode 100644 index 48b8f593..00000000 --- a/.github/ISSUE_TEMPLATE/feature_request.md +++ /dev/null @@ -1,32 +0,0 @@ ---- -name: Feature request -about: Suggest an idea for the nf-core/mag pipeline -labels: enhancement ---- - - - -## Is your feature request related to a problem? Please describe - - - - - -## Describe the solution you'd like - - - -## Describe alternatives you've considered - - - -## Additional context - - diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml new file mode 100644 index 00000000..7fa13dca --- /dev/null +++ b/.github/ISSUE_TEMPLATE/feature_request.yml @@ -0,0 +1,11 @@ +name: Feature request +description: Suggest an idea for the nf-core/mag pipeline +labels: enhancement +body: + - type: textarea + id: description + attributes: + label: Description of feature + description: Please describe your suggestion for a new feature. It might help to describe a problem or use case, plus any alternatives that you have considered. + validations: + required: true diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 9d3d0ca5..150acfbf 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -14,14 +14,14 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@master + uses: nf-core/tower-action@v2 # TODO nf-core: You can customise AWS full pipeline tests as required # Add full size test data (but still relatively small datasets for few samples) # on the `test_full.config` test runs with only one set of parameters with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} - bearer_token: ${{ secrets.TOWER_BEARER_TOKEN }} + access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} pipeline: ${{ github.repository }} revision: ${{ github.sha }} @@ -30,5 +30,5 @@ jobs: { "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-${{ github.sha }}" } - profiles: '[ "test_full", "aws_tower" ]' - + profiles: test_full,aws_tower + pre_run_script: 'export NXF_VER=21.10.3' diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 4e3797f9..f42d2878 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -11,18 +11,18 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@master + uses: nf-core/tower-action@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} - bearer_token: ${{ secrets.TOWER_BEARER_TOKEN }} + access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} pipeline: ${{ github.repository }} revision: ${{ github.sha }} workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/mag/work-${{ github.sha }} parameters: | { - "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-${{ github.sha }}" + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-test-${{ github.sha }}" } - profiles: '[ "test", "aws_tower" ]' - + profiles: test,aws_tower + pre_run_script: 'export NXF_VER=21.10.3' diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index dad70a3c..59ec52ae 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -8,8 +8,9 @@ on: release: types: [published] -# Uncomment if we need an edge release of Nextflow again -# env: NXF_EDGE: 1 +env: + NXF_ANSI_LOG: false + CAPSULE_LOG: none jobs: test: @@ -17,20 +18,26 @@ jobs: # Only run on push if this is the nf-core dev branch (merged PRs) if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/mag') }} runs-on: ubuntu-latest - env: - NXF_VER: ${{ matrix.nxf_ver }} - NXF_ANSI_LOG: false strategy: matrix: - # Nextflow versions: check pipeline minimum and current latest - nxf_ver: ['21.04.0', ''] + # Nextflow versions + include: + # Test pipeline minimum Nextflow version + - NXF_VER: '21.10.3' + NXF_EDGE: '' + # Test latest edge release of Nextflow + - NXF_VER: '' + NXF_EDGE: '1' steps: - name: Check out pipeline code uses: actions/checkout@v2 - name: Install Nextflow env: - CAPSULE_LOG: none + NXF_VER: ${{ matrix.NXF_VER }} + # Uncomment only if the edge release is more recent than the latest stable release + # See https://github.com/nextflow-io/nextflow/issues/2467 + # NXF_EDGE: ${{ matrix.NXF_EDGE }} run: | wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml index 90f03c6f..44d72994 100644 --- a/.github/workflows/linting_comment.yml +++ b/.github/workflows/linting_comment.yml @@ -15,6 +15,7 @@ jobs: uses: dawidd6/action-download-artifact@v2 with: workflow: linting.yml + workflow_conclusion: completed - name: Get PR number id: pr_number diff --git a/CHANGELOG.md b/CHANGELOG.md index f08beeb9..2854a2b3 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v2.1.0 - [date] +## v2.1.1 - [date] Initial release of nf-core/mag, created with the [nf-core](https://nf-co.re/) template. diff --git a/CITATIONS.md b/CITATIONS.md index ed37431a..256b0739 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -12,7 +12,7 @@ * [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) -* [MultiQC](https://www.ncbi.nlm.nih.gov/pubmed/27312411/) +* [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. ## Software packaging/containerisation tools diff --git a/README.md b/README.md index eac88ee9..7cbbd4d4 100644 --- a/README.md +++ b/README.md @@ -1,11 +1,11 @@ -# ![nf-core/mag](docs/images/nf-core-mag_logo.png) +# ![nf-core/mag](docs/images/nf-core-mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core-mag_logo_dark.png#gh-dark-mode-only) [![GitHub Actions CI Status](https://github.com/nf-core/mag/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) [![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/mag/results) [![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) -[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.04.0-23aa62.svg?labelColor=000000)](https://www.nextflow.io/) +[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.10.3-23aa62.svg?labelColor=000000)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) [![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) [![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) @@ -33,18 +33,21 @@ On release, automated continuous integration tests run the pipeline on a full-si ## Quick Start -1. Install [`Nextflow`](https://www.nextflow.io/docs/latest/getstarted.html#installation) (`>=21.04.0`) +1. Install [`Nextflow`](https://www.nextflow.io/docs/latest/getstarted.html#installation) (`>=21.10.3`) 2. Install any of [`Docker`](https://docs.docker.com/engine/installation/), [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/), [`Podman`](https://podman.io/), [`Shifter`](https://nersc.gitlab.io/development/shifter/how-to-use/) or [`Charliecloud`](https://hpc.github.io/charliecloud/) for full pipeline reproducibility _(please only use [`Conda`](https://conda.io/miniconda.html) as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_ 3. Download the pipeline and test it on a minimal dataset with a single command: ```console - nextflow run nf-core/mag -profile test, + nextflow run nf-core/mag -profile test,YOURPROFILE ``` + Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. + + > * The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. > * Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. - > * If you are using `singularity` then the pipeline will auto-detect this and attempt to download the Singularity images directly as opposed to performing a conversion from Docker images. If you are persistently observing issues downloading Singularity images directly due to timeout or network issues then please use the `--singularity_pull_docker_container` parameter to pull and convert the Docker image instead. Alternatively, it is highly recommended to use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to pre-download all of the required containers before running the pipeline and to set the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options to be able to store and re-use the images from a central location for future pipeline runs. + > * If you are using `singularity` and are persistently observing issues downloading Singularity images directly due to timeout or network issues, then you can use the `--singularity_pull_docker_container` parameter to pull and convert the Docker image instead. Alternatively, you can use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. > * If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. 4. Start running your own analysis! diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml index 5bacff3f..7b6b9073 100644 --- a/assets/multiqc_config.yaml +++ b/assets/multiqc_config.yaml @@ -1,7 +1,7 @@ report_comment: > This report has been generated by the nf-core/mag analysis pipeline. 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Resource Limit Is Reached

+The website is temporarily unable to service your request as it exceeded resource limit. +Please try again later. + diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt index faf9a292..34191548 100644 --- a/assets/sendmail_template.txt +++ b/assets/sendmail_template.txt @@ -12,9 +12,9 @@ $email_html Content-Type: image/png;name="nf-core-mag_logo.png" Content-Transfer-Encoding: base64 Content-ID: -Content-Disposition: inline; filename="nf-core-mag_logo.png" +Content-Disposition: inline; filename="nf-core-mag_logo_light.png" -<% out << new File("$projectDir/assets/nf-core-mag_logo.png"). +<% out << new File("$projectDir/assets/nf-core-mag_logo_light.png"). bytes. encodeBase64(). toString(). diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py deleted file mode 100755 index 2da103e8..00000000 --- a/bin/scrape_software_versions.py +++ /dev/null @@ -1,36 +0,0 @@ -#!/usr/bin/env python -from __future__ import print_function -import os - -results = {} -version_files = [x for x in os.listdir(".") if x.endswith(".version.txt")] -for version_file in version_files: - - software = version_file.replace(".version.txt", "") - if software == "pipeline": - software = "nf-core/mag" - - with open(version_file) as fin: - version = fin.read().strip() - results[software] = version - -# Dump to YAML -print( - """ -id: 'software_versions' -section_name: 'nf-core/mag Software Versions' -section_href: 'https://github.com/nf-core/mag' -plot_type: 'html' -description: 'are collected at run time from the software output.' -data: | -
-""" -) -for k, v in sorted(results.items()): - print("
{}
{}
".format(k, v)) -print("
") - -# Write out as tsv file: -with open("software_versions.tsv", "w") as f: - for k, v in sorted(results.items()): - f.write("{}\t{}\n".format(k, v)) diff --git a/conf/base.config b/conf/base.config index 34300497..8ee679ad 100644 --- a/conf/base.config +++ b/conf/base.config @@ -54,4 +54,7 @@ process { errorStrategy = 'retry' maxRetries = 2 } + withName:CUSTOM_DUMPSOFTWAREVERSIONS { + cache = false + } } diff --git a/conf/modules.config b/conf/modules.config index 0b1bfdec..a0506a4d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -1,32 +1,41 @@ /* ======================================================================================== - Config file for defining DSL2 per module options + Config file for defining DSL2 per module options and publishing paths ======================================================================================== Available keys to override module options: - args = Additional arguments appended to command in module. - args2 = Second set of arguments appended to command in module (multi-tool modules). - args3 = Third set of arguments appended to command in module (multi-tool modules). - publish_dir = Directory to publish results. - publish_by_meta = Groovy list of keys available in meta map to append as directories to "publish_dir" path - If publish_by_meta = true - Value of ${meta['id']} is appended as a directory to "publish_dir" path - If publish_by_meta = ['id', 'custompath'] - If "id" is in meta map and "custompath" isn't then "${meta['id']}/custompath/" - is appended as a directory to "publish_dir" path - If publish_by_meta = false / null - No directories are appended to "publish_dir" path - publish_files = Groovy map where key = "file_ext" and value = "directory" to publish results for that file extension - The value of "directory" is appended to the standard "publish_dir" path as defined above. - If publish_files = null (unspecified) - All files are published. - If publish_files = false - No files are published. - suffix = File name suffix for output files. + ext.args = Additional arguments appended to command in module. + ext.args2 = Second set of arguments appended to command in module (multi-tool modules). + ext.args3 = Third set of arguments appended to command in module (multi-tool modules). + ext.prefix = File name prefix for output files. ---------------------------------------------------------------------------------------- */ -params { - modules { - 'fastqc' { - args = "--quiet" - } - 'multiqc' { - args = "" - } +process { + + publishDir = [ + path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + mode: 'copy', + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + + withName: SAMPLESHEET_CHECK { + publishDir = [ + path: { "${params.outdir}/pipeline_info" }, + mode: 'copy', + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + + withName: FASTQC { + ext.args = '--quiet' } + + withName: CUSTOM_DUMPSOFTWAREVERSIONS { + publishDir = [ + path: { "${params.outdir}/pipeline_info" }, + mode: 'copy', + pattern: '*_versions.yml' + ] + } + } diff --git a/conf/test.config b/conf/test.config index 8e39d50b..6ce1192d 100644 --- a/conf/test.config +++ b/conf/test.config @@ -16,8 +16,8 @@ params { // Limit resources so that this can run on GitHub Actions max_cpus = 2 - max_memory = 6.GB - max_time = 6.h + max_memory = '6.GB' + max_time = '6.h' // Input data // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets diff --git a/docs/images/nf-core-mag_logo.png b/docs/images/nf-core-mag_logo.png deleted file mode 100644 index 628fde968941e85f8120121577570c8709f45901..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 30598 zcmYhi1yodBxIa9A(lJPPiL@XcLze>5Eezcq!qBOdfRfTB-QCjNT}ls)bk}!y@4f%^ zorTM_#EG-dexB!7J4{(o77Lve9RvbleRwaW3IZW&gFpx`P?3RuIq#Gk0scaBe6QmI 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Service Unavailable

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The server is temporarily unable to service your +request due to maintenance downtime or capacity +problems. Please try again later.

+

Additionally, a 503 Service Unavailable +error was encountered while trying to use an ErrorDocument to handle the request.

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z_0ZArZM?0Z9je4;_Go$t3imKQsM>MQEm$0mGNY-p%n7|L3QvQpHt2)I@kogvBZ$%_ zKHvlWYX(Bqkvl^5Kt5FMT@tZuMZ?q(Rbj?>6$lZR@I#Q9k9Z6XCr#3W@<^mrv- zxOp#HEVNy58?@M3^j$o^8RhSMK4pcLqbCVPS@h9Y+B~uft&{ia*>)2}P1;(6Hx$Bf znpUVgA#gs+e!}yAUX!ucz=EPeaojLxxRhMBjap*6?r?b-?QIA76*@nP2bDHV4Pf&80`n4Gdj#`5VEZe*v2U(}e&4 literal 0 HcmV?d00001 diff --git a/docs/output.md b/docs/output.md index 7aaa57fb..a2782c16 100644 --- a/docs/output.md +++ b/docs/output.md @@ -60,7 +60,7 @@ Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQ * `pipeline_info/` * Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. - * Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.tsv`. + * Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. * Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`. diff --git a/docs/usage.md b/docs/usage.md index 7770d250..db7f656b 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -181,42 +181,6 @@ process { > **NB:** We specify just the process name i.e. `STAR_ALIGN` in the config file and not the full task name string that is printed to screen in the error message or on the terminal whilst the pipeline is running i.e. `RNASEQ:ALIGN_STAR:STAR_ALIGN`. You may get a warning suggesting that the process selector isn't recognised but you can ignore that if the process name has been specified correctly. This is something that needs to be fixed upstream in core Nextflow. -### Tool-specific options - -For the ultimate flexibility, we have implemented and are using Nextflow DSL2 modules in a way where it is possible for both developers and users to change tool-specific command-line arguments (e.g. providing an additional command-line argument to the `STAR_ALIGN` process) as well as publishing options (e.g. saving files produced by the `STAR_ALIGN` process that aren't saved by default by the pipeline). In the majority of instances, as a user you won't have to change the default options set by the pipeline developer(s), however, there may be edge cases where creating a simple custom config file can improve the behaviour of the pipeline if for example it is failing due to a weird error that requires setting a tool-specific parameter to deal with smaller / larger genomes. - -The command-line arguments passed to STAR in the `STAR_ALIGN` module are a combination of: - -* Mandatory arguments or those that need to be evaluated within the scope of the module, as supplied in the [`script`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L49-L55) section of the module file. - -* An [`options.args`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L56) string of non-mandatory parameters that is set to be empty by default in the module but can be overwritten when including the module in the sub-workflow / workflow context via the `addParams` Nextflow option. - -The nf-core/rnaseq pipeline has a sub-workflow (see [terminology](https://github.com/nf-core/modules#terminology)) specifically to align reads with STAR and to sort, index and generate some basic stats on the resulting BAM files using SAMtools. At the top of this file we import the `STAR_ALIGN` module via the Nextflow [`include`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/subworkflows/nf-core/align_star.nf#L10) keyword and by default the options passed to the module via the `addParams` option are set as an empty Groovy map [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/subworkflows/nf-core/align_star.nf#L5); this in turn means `options.args` will be set to empty by default in the module file too. This is an intentional design choice and allows us to implement well-written sub-workflows composed of a chain of tools that by default run with the bare minimum parameter set for any given tool in order to make it much easier to share across pipelines and to provide the flexibility for users and developers to customise any non-mandatory arguments. - -When including the sub-workflow above in the main pipeline workflow we use the same `include` statement, however, we now have the ability to overwrite options for each of the tools in the sub-workflow including the [`align_options`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/workflows/rnaseq.nf#L225) variable that will be used specifically to overwrite the optional arguments passed to the `STAR_ALIGN` module. In this case, the options to be provided to `STAR_ALIGN` have been assigned sensible defaults by the developer(s) in the pipeline's [`modules.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/modules.config#L70-L74) and can be accessed and customised in the [workflow context](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/workflows/rnaseq.nf#L201-L204) too before eventually passing them to the sub-workflow as a Groovy map called `star_align_options`. These options will then be propagated from `workflow -> sub-workflow -> module`. - -As mentioned at the beginning of this section it may also be necessary for users to overwrite the options passed to modules to be able to customise specific aspects of the way in which a particular tool is executed by the pipeline. Given that all of the default module options are stored in the pipeline's `modules.config` as a [`params` variable](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/modules.config#L24-L25) it is also possible to overwrite any of these options via a custom config file. - -Say for example we want to append an additional, non-mandatory parameter (i.e. `--outFilterMismatchNmax 16`) to the arguments passed to the `STAR_ALIGN` module. Firstly, we need to copy across the default `args` specified in the [`modules.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/modules.config#L71) and create a custom config file that is a composite of the default `args` as well as the additional options you would like to provide. This is very important because Nextflow will overwrite the default value of `args` that you provide via the custom config. - -As you will see in the example below, we have: - -* appended `--outFilterMismatchNmax 16` to the default `args` used by the module. -* changed the default `publish_dir` value to where the files will eventually be published in the main results directory. -* appended `'bam':''` to the default value of `publish_files` so that the BAM files generated by the process will also be saved in the top-level results directory for the module. Note: `'out':'log'` means any file/directory ending in `out` will now be saved in a separate directory called `my_star_directory/log/`. - -```nextflow -params { - modules { - 'star_align' { - args = "--quantMode TranscriptomeSAM --twopassMode Basic --outSAMtype BAM Unsorted --readFilesCommand zcat --runRNGseed 0 --outFilterMultimapNmax 20 --alignSJDBoverhangMin 1 --outSAMattributes NH HI AS NM MD --quantTranscriptomeBan Singleend --outFilterMismatchNmax 16" - publish_dir = "my_star_directory" - publish_files = ['out':'log', 'tab':'log', 'bam':''] - } - } -} -``` - ### Updating containers The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. If for some reason you need to use a different version of a particular tool with the pipeline then you just need to identify the `process` name and override the Nextflow `container` definition for that process using the `withName` declaration. For example, in the [nf-core/viralrecon](https://nf-co.re/viralrecon) pipeline a tool called [Pangolin](https://github.com/cov-lineages/pangolin) has been used during the COVID-19 pandemic to assign lineages to SARS-CoV-2 genome sequenced samples. Given that the lineage assignments change quite frequently it doesn't make sense to re-release the nf-core/viralrecon everytime a new version of Pangolin has been released. However, you can override the default container used by the pipeline by creating a custom config file and passing it as a command-line argument via `-c custom.config`. diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy index 8d6920dd..40ab65f2 100755 --- a/lib/NfcoreSchema.groovy +++ b/lib/NfcoreSchema.groovy @@ -105,9 +105,13 @@ class NfcoreSchema { // Collect expected parameters from the schema def expectedParams = [] + def enums = [:] for (group in schemaParams) { for (p in group.value['properties']) { expectedParams.push(p.key) + if (group.value['properties'][p.key].containsKey('enum')) { + enums[p.key] = group.value['properties'][p.key]['enum'] + } } } @@ -155,7 +159,7 @@ class NfcoreSchema { println '' log.error 'ERROR: Validation of pipeline parameters failed!' JSONObject exceptionJSON = e.toJSON() - printExceptions(exceptionJSON, params_json, log) + printExceptions(exceptionJSON, params_json, log, enums) println '' has_error = true } @@ -202,7 +206,7 @@ class NfcoreSchema { } def type = '[' + group_params.get(param).type + ']' def description = group_params.get(param).description - def defaultValue = group_params.get(param).default ? " [default: " + group_params.get(param).default.toString() + "]" : '' + def defaultValue = group_params.get(param).default != null ? " [default: " + group_params.get(param).default.toString() + "]" : '' def description_default = description + colors.dim + defaultValue + colors.reset // Wrap long description texts // Loosely based on https://dzone.com/articles/groovy-plain-text-word-wrap @@ -260,13 +264,12 @@ class NfcoreSchema { // Get pipeline parameters defined in JSON Schema def Map params_summary = [:] - def blacklist = ['hostnames'] def params_map = paramsLoad(getSchemaPath(workflow, schema_filename=schema_filename)) for (group in params_map.keySet()) { def sub_params = new LinkedHashMap() def group_params = params_map.get(group) // This gets the parameters of that particular group for (param in group_params.keySet()) { - if (params.containsKey(param) && !blacklist.contains(param)) { + if (params.containsKey(param)) { def params_value = params.get(param) def schema_value = group_params.get(param).default def param_type = group_params.get(param).type @@ -330,7 +333,7 @@ class NfcoreSchema { // // Loop over nested exceptions and print the causingException // - private static void printExceptions(ex_json, params_json, log) { + private static void printExceptions(ex_json, params_json, log, enums, limit=5) { def causingExceptions = ex_json['causingExceptions'] if (causingExceptions.length() == 0) { def m = ex_json['message'] =~ /required key \[([^\]]+)\] not found/ @@ -346,11 +349,20 @@ class NfcoreSchema { else { def param = ex_json['pointerToViolation'] - ~/^#\// def param_val = params_json[param].toString() - log.error "* --${param}: ${ex_json['message']} (${param_val})" + if (enums.containsKey(param)) { + def error_msg = "* --${param}: '${param_val}' is not a valid choice (Available choices" + if (enums[param].size() > limit) { + log.error "${error_msg} (${limit} of ${enums[param].size()}): ${enums[param][0..limit-1].join(', ')}, ... )" + } else { + log.error "${error_msg}: ${enums[param].join(', ')})" + } + } else { + log.error "* --${param}: ${ex_json['message']} (${param_val})" + } } } for (ex in causingExceptions) { - printExceptions(ex, params_json, log) + printExceptions(ex, params_json, log, enums) } } diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 44551e0a..2fc0a9b9 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -19,27 +19,16 @@ class NfcoreTemplate { } // - // Check params.hostnames + // Warn if a -profile or Nextflow config has not been provided to run the pipeline // - public static void hostName(workflow, params, log) { - Map colors = logColours(params.monochrome_logs) - if (params.hostnames) { - try { - def hostname = "hostname".execute().text.trim() - params.hostnames.each { prof, hnames -> - hnames.each { hname -> - if (hostname.contains(hname) && !workflow.profile.contains(prof)) { - log.info "=${colors.yellow}====================================================${colors.reset}=\n" + - "${colors.yellow}WARN: You are running with `-profile $workflow.profile`\n" + - " but your machine hostname is ${colors.white}'$hostname'${colors.reset}.\n" + - " ${colors.yellow_bold}Please use `-profile $prof${colors.reset}`\n" + - "=${colors.yellow}====================================================${colors.reset}=" - } - } - } - } catch (Exception e) { - log.warn "[$workflow.manifest.name] Could not determine 'hostname' - skipping check. Reason: ${e.message}." - } + public static void checkConfigProvided(workflow, log) { + if (workflow.profile == 'standard' && workflow.configFiles.size() <= 1) { + log.warn "[$workflow.manifest.name] You are attempting to run the pipeline without any custom configuration!\n\n" + + "This will be dependent on your local compute environment but can be achieved via one or more of the following:\n" + + " (1) Using an existing pipeline profile e.g. `-profile docker` or `-profile singularity`\n" + + " (2) Using an existing nf-core/configs for your Institution e.g. `-profile crick` or `-profile uppmax`\n" + + " (3) Using your own local custom config e.g. `-c /path/to/your/custom.config`\n\n" + + "Please refer to the quick start section and usage docs for the pipeline.\n " } } @@ -168,7 +157,6 @@ class NfcoreTemplate { log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed successfully, but with errored process(es) ${colors.reset}-" } } else { - hostName(workflow, params, log) log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed with errors${colors.reset}-" } } diff --git a/lib/Utils.groovy b/lib/Utils.groovy index 18173e98..1b88aec0 100755 --- a/lib/Utils.groovy +++ b/lib/Utils.groovy @@ -37,11 +37,4 @@ class Utils { "===================================================================================" } } - - // - // Join module args with appropriate spacing - // - public static String joinModuleArgs(args_list) { - return ' ' + args_list.join(' ') - } } diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index 369fee93..317c61e2 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -61,6 +61,9 @@ class WorkflowMain { // Print parameter summary log to screen log.info paramsSummaryLog(workflow, params, log) + // Check that a -profile or Nextflow config has been provided to run the pipeline + NfcoreTemplate.checkConfigProvided(workflow, log) + // Check that conda channels are set-up correctly if (params.enable_conda) { Utils.checkCondaChannels(log) @@ -69,9 +72,6 @@ class WorkflowMain { // Check AWS batch settings NfcoreTemplate.awsBatch(workflow, params) - // Check the hostnames against configured profiles - NfcoreTemplate.hostName(workflow, params, log) - // Check input has been provided if (!params.input) { log.error "Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'" diff --git a/modules.json b/modules.json index 45719b4e..6394e068 100644 --- a/modules.json +++ b/modules.json @@ -3,12 +3,15 @@ "homePage": "https://github.com/nf-core/mag", "repos": { "nf-core/modules": { + "custom/dumpsoftwareversions": { + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + }, "fastqc": { - "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" + "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "multiqc": { - "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" + "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" } } } -} +} \ No newline at end of file diff --git a/modules/local/functions.nf b/modules/local/functions.nf deleted file mode 100644 index da9da093..00000000 --- a/modules/local/functions.nf +++ /dev/null @@ -1,68 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - if (!args.filename.endsWith('.version.txt')) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } - } -} diff --git a/modules/local/get_software_versions.nf b/modules/local/get_software_versions.nf deleted file mode 100644 index 15886433..00000000 --- a/modules/local/get_software_versions.nf +++ /dev/null @@ -1,33 +0,0 @@ -// Import generic module functions -include { saveFiles } from './functions' - -params.options = [:] - -process GET_SOFTWARE_VERSIONS { - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:'pipeline_info', meta:[:], publish_by_meta:[]) } - - conda (params.enable_conda ? "conda-forge::python=3.8.3" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/python:3.8.3" - } else { - container "quay.io/biocontainers/python:3.8.3" - } - - cache false - - input: - path versions - - output: - path "software_versions.tsv" , emit: tsv - path 'software_versions_mqc.yaml', emit: yaml - - script: // This script is bundled with the pipeline, in nf-core/mag/bin/ - """ - echo $workflow.manifest.version > pipeline.version.txt - echo $workflow.nextflow.version > nextflow.version.txt - scrape_software_versions.py &> software_versions_mqc.yaml - """ -} diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf index cf0a3c8d..aa7269ce 100644 --- a/modules/local/samplesheet_check.nf +++ b/modules/local/samplesheet_check.nf @@ -1,31 +1,27 @@ -// Import generic module functions -include { saveFiles } from './functions' - -params.options = [:] - process SAMPLESHEET_CHECK { tag "$samplesheet" - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:'pipeline_info', meta:[:], publish_by_meta:[]) } conda (params.enable_conda ? "conda-forge::python=3.8.3" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/python:3.8.3" - } else { - container "quay.io/biocontainers/python:3.8.3" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/python:3.8.3' : + 'quay.io/biocontainers/python:3.8.3' }" input: path samplesheet output: - path '*.csv' + path '*.csv' , emit: csv + path "versions.yml", emit: versions script: // This script is bundled with the pipeline, in nf-core/mag/bin/ """ check_samplesheet.py \\ $samplesheet \\ samplesheet.valid.csv + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + python: \$(python --version | sed 's/Python //g') + END_VERSIONS """ } diff --git a/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf b/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf new file mode 100644 index 00000000..934bb467 --- /dev/null +++ b/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf @@ -0,0 +1,21 @@ +process CUSTOM_DUMPSOFTWAREVERSIONS { + label 'process_low' + + // Requires `pyyaml` which does not have a dedicated container but is in the MultiQC container + conda (params.enable_conda ? "bioconda::multiqc=1.11" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.11--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.11--pyhdfd78af_0' }" + + input: + path versions + + output: + path "software_versions.yml" , emit: yml + path "software_versions_mqc.yml", emit: mqc_yml + path "versions.yml" , emit: versions + + script: + def args = task.ext.args ?: '' + template 'dumpsoftwareversions.py' +} diff --git a/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml new file mode 100644 index 00000000..5b5b8a60 --- /dev/null +++ b/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml @@ -0,0 +1,34 @@ +name: custom_dumpsoftwareversions +description: Custom module used to dump software versions within the nf-core pipeline template +keywords: + - custom + - version +tools: + - custom: + description: Custom module used to dump software versions within the nf-core pipeline template + homepage: https://github.com/nf-core/tools + documentation: https://github.com/nf-core/tools + licence: ['MIT'] +input: + - versions: + type: file + description: YML file containing software versions + pattern: "*.yml" + +output: + - yml: + type: file + description: Standard YML file containing software versions + pattern: "software_versions.yml" + - mqc_yml: + type: file + description: MultiQC custom content YML file containing software versions + pattern: "software_versions_mqc.yml" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + +authors: + - "@drpatelh" + - "@grst" diff --git a/modules/nf-core/modules/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py b/modules/nf-core/modules/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py new file mode 100644 index 00000000..d1390392 --- /dev/null +++ b/modules/nf-core/modules/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py @@ -0,0 +1,89 @@ +#!/usr/bin/env python + +import yaml +import platform +from textwrap import dedent + + +def _make_versions_html(versions): + html = [ + dedent( + """\\ + + + + + + + + + + """ + ) + ] + for process, tmp_versions in sorted(versions.items()): + html.append("") + for i, (tool, version) in enumerate(sorted(tmp_versions.items())): + html.append( + dedent( + f"""\\ + + + + + + """ + ) + ) + html.append("") + html.append("
Process Name Software Version
{process if (i == 0) else ''}{tool}{version}
") + return "\\n".join(html) + + +versions_this_module = {} +versions_this_module["${task.process}"] = { + "python": platform.python_version(), + "yaml": yaml.__version__, +} + +with open("$versions") as f: + versions_by_process = yaml.load(f, Loader=yaml.BaseLoader) | versions_this_module + +# aggregate versions by the module name (derived from fully-qualified process name) +versions_by_module = {} +for process, process_versions in versions_by_process.items(): + module = process.split(":")[-1] + try: + assert versions_by_module[module] == process_versions, ( + "We assume that software versions are the same between all modules. " + "If you see this error-message it means you discovered an edge-case " + "and should open an issue in nf-core/tools. " + ) + except KeyError: + versions_by_module[module] = process_versions + +versions_by_module["Workflow"] = { + "Nextflow": "$workflow.nextflow.version", + "$workflow.manifest.name": "$workflow.manifest.version", +} + +versions_mqc = { + "id": "software_versions", + "section_name": "${workflow.manifest.name} Software Versions", + "section_href": "https://github.com/${workflow.manifest.name}", + "plot_type": "html", + "description": "are collected at run time from the software output.", + "data": _make_versions_html(versions_by_module), +} + +with open("software_versions.yml", "w") as f: + yaml.dump(versions_by_module, f, default_flow_style=False) +with open("software_versions_mqc.yml", "w") as f: + yaml.dump(versions_mqc, f, default_flow_style=False) + +with open("versions.yml", "w") as f: + yaml.dump(versions_this_module, f, default_flow_style=False) diff --git a/modules/nf-core/modules/fastqc/functions.nf b/modules/nf-core/modules/fastqc/functions.nf deleted file mode 100644 index da9da093..00000000 --- a/modules/nf-core/modules/fastqc/functions.nf +++ /dev/null @@ -1,68 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - if (!args.filename.endsWith('.version.txt')) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } - } -} diff --git a/modules/nf-core/modules/fastqc/main.nf b/modules/nf-core/modules/fastqc/main.nf index 39c327b2..d250eca0 100644 --- a/modules/nf-core/modules/fastqc/main.nf +++ b/modules/nf-core/modules/fastqc/main.nf @@ -1,22 +1,11 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process FASTQC { tag "$meta.id" label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['id']) } conda (params.enable_conda ? "bioconda::fastqc=0.11.9" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0" - } else { - container "quay.io/biocontainers/fastqc:0.11.9--0" - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0' : + 'quay.io/biocontainers/fastqc:0.11.9--0' }" input: tuple val(meta), path(reads) @@ -24,24 +13,32 @@ process FASTQC { output: tuple val(meta), path("*.html"), emit: html tuple val(meta), path("*.zip") , emit: zip - path "*.version.txt" , emit: version + path "versions.yml" , emit: versions script: + def args = task.ext.args ?: '' // Add soft-links to original FastQs for consistent naming in pipeline - def software = getSoftwareName(task.process) - def prefix = options.suffix ? "${meta.id}${options.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" if (meta.single_end) { """ [ ! -f ${prefix}.fastq.gz ] && ln -s $reads ${prefix}.fastq.gz - fastqc $options.args --threads $task.cpus ${prefix}.fastq.gz - fastqc --version | sed -e "s/FastQC v//g" > ${software}.version.txt + fastqc $args --threads $task.cpus ${prefix}.fastq.gz + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" ) + END_VERSIONS """ } else { """ [ ! -f ${prefix}_1.fastq.gz ] && ln -s ${reads[0]} ${prefix}_1.fastq.gz [ ! -f ${prefix}_2.fastq.gz ] && ln -s ${reads[1]} ${prefix}_2.fastq.gz - fastqc $options.args --threads $task.cpus ${prefix}_1.fastq.gz ${prefix}_2.fastq.gz - fastqc --version | sed -e "s/FastQC v//g" > ${software}.version.txt + fastqc $args --threads $task.cpus ${prefix}_1.fastq.gz ${prefix}_2.fastq.gz + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" ) + END_VERSIONS """ } } diff --git a/modules/nf-core/modules/fastqc/meta.yml b/modules/nf-core/modules/fastqc/meta.yml index 8eb9953d..b09553a3 100644 --- a/modules/nf-core/modules/fastqc/meta.yml +++ b/modules/nf-core/modules/fastqc/meta.yml @@ -15,6 +15,7 @@ tools: overrepresented sequences. homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ + licence: ['GPL-2.0-only'] input: - meta: type: map @@ -40,10 +41,10 @@ output: type: file description: FastQC report archive pattern: "*_{fastqc.zip}" - - version: + - versions: type: file - description: File containing software version - pattern: "*.{version.txt}" + description: File containing software versions + pattern: "versions.yml" authors: - "@drpatelh" - "@grst" diff --git a/modules/nf-core/modules/multiqc/functions.nf b/modules/nf-core/modules/multiqc/functions.nf deleted file mode 100644 index da9da093..00000000 --- a/modules/nf-core/modules/multiqc/functions.nf +++ /dev/null @@ -1,68 +0,0 @@ -// -// Utility functions used in nf-core DSL2 module files -// - -// -// Extract name of software tool from process name using $task.process -// -def getSoftwareName(task_process) { - return task_process.tokenize(':')[-1].tokenize('_')[0].toLowerCase() -} - -// -// Function to initialise default values and to generate a Groovy Map of available options for nf-core modules -// -def initOptions(Map args) { - def Map options = [:] - options.args = args.args ?: '' - options.args2 = args.args2 ?: '' - options.args3 = args.args3 ?: '' - options.publish_by_meta = args.publish_by_meta ?: [] - options.publish_dir = args.publish_dir ?: '' - options.publish_files = args.publish_files - options.suffix = args.suffix ?: '' - return options -} - -// -// Tidy up and join elements of a list to return a path string -// -def getPathFromList(path_list) { - def paths = path_list.findAll { item -> !item?.trim().isEmpty() } // Remove empty entries - paths = paths.collect { it.trim().replaceAll("^[/]+|[/]+\$", "") } // Trim whitespace and trailing slashes - return paths.join('/') -} - -// -// Function to save/publish module results -// -def saveFiles(Map args) { - if (!args.filename.endsWith('.version.txt')) { - def ioptions = initOptions(args.options) - def path_list = [ ioptions.publish_dir ?: args.publish_dir ] - if (ioptions.publish_by_meta) { - def key_list = ioptions.publish_by_meta instanceof List ? ioptions.publish_by_meta : args.publish_by_meta - for (key in key_list) { - if (args.meta && key instanceof String) { - def path = key - if (args.meta.containsKey(key)) { - path = args.meta[key] instanceof Boolean ? "${key}_${args.meta[key]}".toString() : args.meta[key] - } - path = path instanceof String ? path : '' - path_list.add(path) - } - } - } - if (ioptions.publish_files instanceof Map) { - for (ext in ioptions.publish_files) { - if (args.filename.endsWith(ext.key)) { - def ext_list = path_list.collect() - ext_list.add(ext.value) - return "${getPathFromList(ext_list)}/$args.filename" - } - } - } else if (ioptions.publish_files == null) { - return "${getPathFromList(path_list)}/$args.filename" - } - } -} diff --git a/modules/nf-core/modules/multiqc/main.nf b/modules/nf-core/modules/multiqc/main.nf index da780800..3dceb162 100644 --- a/modules/nf-core/modules/multiqc/main.nf +++ b/modules/nf-core/modules/multiqc/main.nf @@ -1,21 +1,10 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName } from './functions' - -params.options = [:] -options = initOptions(params.options) - process MULTIQC { label 'process_medium' - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:[:], publish_by_meta:[]) } - conda (params.enable_conda ? "bioconda::multiqc=1.10.1" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/multiqc:1.10.1--py_0" - } else { - container "quay.io/biocontainers/multiqc:1.10.1--py_0" - } + conda (params.enable_conda ? 'bioconda::multiqc=1.11' : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.11--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.11--pyhdfd78af_0' }" input: path multiqc_files @@ -24,12 +13,16 @@ process MULTIQC { path "*multiqc_report.html", emit: report path "*_data" , emit: data path "*_plots" , optional:true, emit: plots - path "*.version.txt" , emit: version + path "versions.yml" , emit: versions script: - def software = getSoftwareName(task.process) + def args = task.ext.args ?: '' """ - multiqc -f $options.args . - multiqc --version | sed -e "s/multiqc, version //g" > ${software}.version.txt + multiqc -f $args . + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" ) + END_VERSIONS """ } diff --git a/modules/nf-core/modules/multiqc/meta.yml b/modules/nf-core/modules/multiqc/meta.yml index 532a8bb1..63c75a45 100644 --- a/modules/nf-core/modules/multiqc/meta.yml +++ b/modules/nf-core/modules/multiqc/meta.yml @@ -11,6 +11,7 @@ tools: It's a general use tool, perfect for summarising the output from numerous bioinformatics tools. homepage: https://multiqc.info/ documentation: https://multiqc.info/docs/ + licence: ['GPL-3.0-or-later'] input: - multiqc_files: type: file @@ -29,10 +30,10 @@ output: type: file description: Plots created by MultiQC pattern: "*_data" - - version: + - versions: type: file - description: File containing software version - pattern: "*.{version.txt}" + description: File containing software versions + pattern: "versions.yml" authors: - "@abhi18av" - "@bunop" diff --git a/nextflow.config b/nextflow.config index 54f0ad43..6a929ee8 100644 --- a/nextflow.config +++ b/nextflow.config @@ -26,7 +26,6 @@ params { // Boilerplate options outdir = './results' tracedir = "${params.outdir}/pipeline_info" - publish_dir_mode = 'copy' email = null email_on_fail = null plaintext_email = false @@ -34,14 +33,12 @@ params { help = false validate_params = true show_hidden_params = false - schema_ignore_params = 'genomes,modules' + schema_ignore_params = 'genomes' enable_conda = false - singularity_pull_docker_container = false // Config options custom_config_version = 'master' custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" - hostnames = [:] config_profile_description = null config_profile_contact = null config_profile_url = null @@ -58,9 +55,6 @@ params { // Load base.config by default for all pipelines includeConfig 'conf/base.config' -// Load modules.config for DSL2 module specific options -includeConfig 'conf/modules.config' - // Load nf-core custom profiles from different Institutions try { includeConfig "${params.custom_config_base}/nfcore_custom.config" @@ -68,13 +62,6 @@ try { System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config") } -// Load igenomes.config if required -if (!params.igenomes_ignore) { - includeConfig 'conf/igenomes.config' -} else { - params.genomes = [:] -} - profiles { debug { process.beforeScript = 'echo $HOSTNAME' } conda { @@ -126,11 +113,22 @@ profiles { test_full { includeConfig 'conf/test_full.config' } } +// Load igenomes.config if required +if (!params.igenomes_ignore) { + includeConfig 'conf/igenomes.config' +} else { + params.genomes = [:] +} + // Export these variables to prevent local Python/R libraries from conflicting with those in the container +// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. +// See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable. + env { PYTHONNOUSERSITE = 1 R_PROFILE_USER = "/.Rprofile" R_ENVIRON_USER = "/.Renviron" + JULIA_DEPOT_PATH = "/usr/local/share/julia" } // Capture exit codes from upstream processes when piping @@ -160,10 +158,13 @@ manifest { homePage = 'https://github.com/nf-core/mag' description = 'Assembly, binning and annotation of metagenomes' mainScript = 'main.nf' - nextflowVersion = '!>=21.04.0' - version = '2.1.0' + nextflowVersion = '!>=21.10.3' + version = '2.1.1' } +// Load modules.config for DSL2 module specific options +includeConfig 'conf/modules.config' + // Function to ensure that resource requirements don't go beyond // a maximum limit def check_max(obj, type) { diff --git a/nextflow_schema.json b/nextflow_schema.json index ae864e42..22133aa1 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -104,12 +104,6 @@ "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", "fa_icon": "fas fa-users-cog" }, - "hostnames": { - "type": "string", - "description": "Institutional configs hostname.", - "hidden": true, - "fa_icon": "fas fa-users-cog" - }, "config_profile_name": { "type": "string", "description": "Institutional config name.", @@ -184,22 +178,6 @@ "fa_icon": "fas fa-question-circle", "hidden": true }, - "publish_dir_mode": { - "type": "string", - "default": "copy", - "description": "Method used to save pipeline results to output directory.", - "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", - "fa_icon": "fas fa-copy", - "enum": [ - "symlink", - "rellink", - "link", - "copy", - "copyNoFollow", - "move" - ], - "hidden": true - }, "email_on_fail": { "type": "string", "description": "Email address for completion summary, only when pipeline fails.", @@ -260,13 +238,6 @@ "description": "Run this workflow with Conda. You can also use '-profile conda' instead of providing this parameter.", "hidden": true, "fa_icon": "fas fa-bacon" - }, - "singularity_pull_docker_container": { - "type": "boolean", - "description": "Instead of directly downloading Singularity images for use with Singularity, force the workflow to pull and convert Docker containers instead.", - "hidden": true, - "fa_icon": "fas fa-toolbox", - "help_text": "This may be useful for example if you are unable to directly pull Singularity containers to run the pipeline due to http/https proxy issues." } } } diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index b664bc8c..cddcbb3c 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -2,9 +2,7 @@ // Check input samplesheet and get read channels // -params.options = [:] - -include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check' addParams( options: params.options ) +include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check' workflow INPUT_CHECK { take: @@ -12,12 +10,14 @@ workflow INPUT_CHECK { main: SAMPLESHEET_CHECK ( samplesheet ) + .csv .splitCsv ( header:true, sep:',' ) .map { create_fastq_channels(it) } .set { reads } emit: - reads // channel: [ val(meta), [ reads ] ] + reads // channel: [ val(meta), [ reads ] ] + versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ] } // Function to get list of [ meta, [ fastq_1, fastq_2 ] ] diff --git a/workflows/mag.nf b/workflows/mag.nf index c712dcba..a454563b 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -32,18 +32,10 @@ ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multi ======================================================================================== */ -// Don't overwrite global params.modules, create a copy instead and use that within the main script. -def modules = params.modules.clone() - -// -// MODULE: Local to the pipeline -// -include { GET_SOFTWARE_VERSIONS } from '../modules/local/get_software_versions' addParams( options: [publish_files : ['tsv':'']] ) - // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' addParams( options: [:] ) +include { INPUT_CHECK } from '../subworkflows/local/input_check' /* ======================================================================================== @@ -51,14 +43,12 @@ include { INPUT_CHECK } from '../subworkflows/local/input_check' addParams( opti ======================================================================================== */ -def multiqc_options = modules['multiqc'] -multiqc_options.args += params.multiqc_title ? Utils.joinModuleArgs(["--title \"$params.multiqc_title\""]) : '' - // // MODULE: Installed directly from nf-core/modules // -include { FASTQC } from '../modules/nf-core/modules/fastqc/main' addParams( options: modules['fastqc'] ) -include { MULTIQC } from '../modules/nf-core/modules/multiqc/main' addParams( options: multiqc_options ) +include { FASTQC } from '../modules/nf-core/modules/fastqc/main' +include { MULTIQC } from '../modules/nf-core/modules/multiqc/main' +include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modules/custom/dumpsoftwareversions/main' /* ======================================================================================== @@ -71,7 +61,7 @@ def multiqc_report = [] workflow MAG { - ch_software_versions = Channel.empty() + ch_versions = Channel.empty() // // SUBWORKFLOW: Read in samplesheet, validate and stage input files @@ -79,6 +69,7 @@ workflow MAG { INPUT_CHECK ( ch_input ) + ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) // // MODULE: Run FastQC @@ -86,21 +77,10 @@ workflow MAG { FASTQC ( INPUT_CHECK.out.reads ) - ch_software_versions = ch_software_versions.mix(FASTQC.out.version.first().ifEmpty(null)) + ch_versions = ch_versions.mix(FASTQC.out.versions.first()) - // - // MODULE: Pipeline reporting - // - ch_software_versions - .map { it -> if (it) [ it.baseName, it ] } - .groupTuple() - .map { it[1][0] } - .flatten() - .collect() - .set { ch_software_versions } - - GET_SOFTWARE_VERSIONS ( - ch_software_versions.map { it }.collect() + CUSTOM_DUMPSOFTWAREVERSIONS ( + ch_versions.unique().collectFile(name: 'collated_versions.yml') ) // @@ -113,14 +93,14 @@ workflow MAG { ch_multiqc_files = ch_multiqc_files.mix(Channel.from(ch_multiqc_config)) ch_multiqc_files = ch_multiqc_files.mix(ch_multiqc_custom_config.collect().ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) - ch_multiqc_files = ch_multiqc_files.mix(GET_SOFTWARE_VERSIONS.out.yaml.collect()) + ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) MULTIQC ( ch_multiqc_files.collect() ) - multiqc_report = MULTIQC.out.report.toList() - ch_software_versions = ch_software_versions.mix(MULTIQC.out.version.ifEmpty(null)) + multiqc_report = MULTIQC.out.report.toList() + ch_versions = ch_versions.mix(MULTIQC.out.versions) } /* From d7e82208a05f0b6803a35960f322fc63a8d3a67c Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 13 Jan 2022 16:22:32 +0100 Subject: [PATCH 032/246] fix: re-add old params inheriance syntax for mag_depths so files published correctly --- conf/modules.config | 5 +++-- workflows/mag.nf | 2 +- 2 files changed, 4 insertions(+), 3 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 5bd7269b..ebe83c17 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -102,13 +102,14 @@ params { publish_dir = "Assembly" } 'mag_depths' { + publish_dir = "GenomeBinning" publish_files = false } 'mag_depths_plot' { - publish_dir = "GenomeBinning/depths" + publish_dir = "GenomeBinning" } 'mag_depths_summary' { - publish_dir = "GenomeBinning/depths" + publish_dir = "GenomeBinning" } 'busco_db_preparation' { publish_files = ['tar.gz':''] diff --git a/workflows/mag.nf b/workflows/mag.nf index d5c75013..d01be81d 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -93,7 +93,7 @@ include { MULTIQC } from '../modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' addParams( bowtie2_align_options: modules['bowtie2_assembly_align'] ) -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' //addParams( metabat2_jgisummarizebamcontigdepths_options: modules['metabat2_jgisummarizecontigdepths'], metabat2_options: modules['metabat2'], mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) +include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' addParams( mag_depths_options: modules['mag_depths'], mag_depths_plot_options: modules['mag_depths_plot'], mag_depths_summary_options: modules['mag_depths_summary']) include { BUSCO_QC } from '../subworkflows/local/busco_qc' addParams( busco_db_options: modules['busco_db_preparation'], busco_options: modules['busco'], busco_save_download_options: modules['busco_save_download'], busco_plot_options: modules['busco_plot'], busco_summary_options: modules['busco_summary']) include { GTDBTK } from '../subworkflows/local/gtdbtk' addParams( gtdbtk_classify_options: modules['gtdbtk_classify'], gtdbtk_summary_options: modules['gtdbtk_summary']) From 634f002d733d4f5f808699270413b3b06116dd05 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 13 Jan 2022 16:45:24 +0100 Subject: [PATCH 033/246] fix: publish jgi depth file with correct pattern --- conf/modules.config | 16 ++++++++-------- subworkflows/local/metabat2_binning.nf | 4 ++-- 2 files changed, 10 insertions(+), 10 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index ebe83c17..704ba28b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -175,19 +175,19 @@ params { // nf-core modules DSL2 syntax v2.0 process { - // publishDir = [ - // path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, - // mode: params.publish_dir_mode, - // saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - //] + // publishDir = [ + // path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + // mode: params.publish_dir_mode, + // saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + //] withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { publishDir = [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2" }, + path: { "${params.outdir}/GenomeBinning/" }, mode: 'copy', - pattern: '*_depth.txt.gz' + pattern: '*-depth.txt.gz' ] - ext.suffix = { "${meta.assembler}-${meta.id}-depth" } + ext.prefix = { "${meta.assembler}-${meta.id}-depth" } } withName: METABAT2_METABAT2 { diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 3de4fe76..62556d78 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -6,9 +6,9 @@ params.mag_depths_options = [:] params.mag_depths_plot_options = [:] params.mag_depths_summary_options = [:] -include { METABAT2_METABAT2 } from '../../modules/nf-core/modules/metabat2/metabat2/main' +include { METABAT2_METABAT2 } from '../../modules/nf-core/modules/metabat2/metabat2/main' include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' -include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' +include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' include { SPLIT_FASTQ } from '../../modules/local/split_fastq' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) From 57380c7bcf2a239fbee459a5241a700455a46f7e Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 13 Jan 2022 20:36:44 +0100 Subject: [PATCH 034/246] fix: split_fasta file publication and name --- conf/modules.config | 2 +- modules/local/{split_fastq.nf => split_fasta.nf} | 4 ++-- subworkflows/local/metabat2_binning.nf | 6 +++--- 3 files changed, 6 insertions(+), 6 deletions(-) rename modules/local/{split_fastq.nf => split_fasta.nf} (94%) diff --git a/conf/modules.config b/conf/modules.config index 704ba28b..10cc734a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -216,7 +216,7 @@ process { ].join(' ').trim() } - withName: SPLIT_FASTQ { + withName: SPLIT_FASTA { publishDir = [ path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, mode: 'copy', diff --git a/modules/local/split_fastq.nf b/modules/local/split_fasta.nf similarity index 94% rename from modules/local/split_fastq.nf rename to modules/local/split_fasta.nf index 20c314a1..73c698d8 100644 --- a/modules/local/split_fastq.nf +++ b/modules/local/split_fasta.nf @@ -1,4 +1,4 @@ -process SPLIT_FASTQ { +process SPLIT_FASTA { tag "${meta.assembler}-${meta.id}" label 'process_low' @@ -14,7 +14,7 @@ process SPLIT_FASTQ { tuple val(meta), path(unbinned) output: - path "unbinned/" , emit: unbinned + path "unbinned/*" , emit: unbinned script: """ diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 62556d78..08efdd05 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -10,7 +10,7 @@ include { METABAT2_METABAT2 } from '../../modules/nf-core/mo include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' -include { SPLIT_FASTQ } from '../../modules/local/split_fastq' +include { SPLIT_FASTA } from '../../modules/local/SPLIT_FASTA' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) @@ -62,7 +62,7 @@ workflow METABAT2_BINNING { // split FASTQ METABAT2_METABAT2.out.unbinned.dump(tag: "input_splitfastq") - SPLIT_FASTQ ( METABAT2_METABAT2.out.unbinned ) + SPLIT_FASTA ( METABAT2_METABAT2.out.unbinned ) // decompress main bins for downstream, have to separate and re-group due to limitation of GUNZIP METABAT2_METABAT2.out.fasta @@ -95,7 +95,7 @@ workflow METABAT2_BINNING { emit: bins = ch_metabat_results_gunzipped - unbinned = SPLIT_FASTQ.out.unbinned + unbinned = SPLIT_FASTA.out.unbinned tooshort = METABAT2_METABAT2.out.tooshort lowdepth = METABAT2_METABAT2.out.lowdepth depths_summary = MAG_DEPTHS_SUMMARY.out.summary From 960c62fcf5b22e25b31af9655a3f3efa1ac438f9 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 14 Jan 2022 07:16:20 +0100 Subject: [PATCH 035/246] fix: wrong include file name --- subworkflows/local/metabat2_binning.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index 08efdd05..cacaca72 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -10,7 +10,7 @@ include { METABAT2_METABAT2 } from '../../modules/nf-core/mo include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' -include { SPLIT_FASTA } from '../../modules/local/SPLIT_FASTA' +include { SPLIT_FASTA } from '../../modules/local/split_fasta' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) From ccd51fe63cae5587827ef72b2a31bf3942b1dddd Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 17 Jan 2022 19:57:37 +0100 Subject: [PATCH 036/246] fix: update split_fastq to ensure can transpose for mixing unbinned with bins for depth plot --- modules/local/split_fasta.nf | 4 +++- subworkflows/local/metabat2_binning.nf | 18 +++++++----------- 2 files changed, 10 insertions(+), 12 deletions(-) diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 73c698d8..93ba6431 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -14,7 +14,9 @@ process SPLIT_FASTA { tuple val(meta), path(unbinned) output: - path "unbinned/*" , emit: unbinned + tuple val(meta), path("unbinned/*unbinned.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do + tuple val(meta), path("unbinned/*unbinned.pooled.fa.gz") , optional:true, emit: pooled + tuple val(meta), path("unbinned/*unbinned.remaining.fa.gz") , optional:true, emit: remaining script: """ diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/metabat2_binning.nf index cacaca72..ec239861 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/metabat2_binning.nf @@ -36,17 +36,14 @@ workflow METABAT2_BINNING { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth - .dump(tag: "output_jgi") .map { it -> it[0]['binner'] = 'METABAT2' [ it[0], it[1] ] } - .dump(tag: "output_jgi_updated") .set { ch_metabat_depths } ch_metabat2_input = assemblies - .dump(tag: "metabat2_raw") .map { it -> it[0]['binner'] = 'METABAT2' @@ -56,26 +53,25 @@ workflow METABAT2_BINNING { .map { it -> [ it[0], it[1], it[4]] } - .dump(tag: "metabat2_input_updated") METABAT2_METABAT2 ( ch_metabat2_input ) // split FASTQ - METABAT2_METABAT2.out.unbinned.dump(tag: "input_splitfastq") + METABAT2_METABAT2.out.unbinned SPLIT_FASTA ( METABAT2_METABAT2.out.unbinned ) - // decompress main bins for downstream, have to separate and re-group due to limitation of GUNZIP - METABAT2_METABAT2.out.fasta - .transpose() - .set { ch_metabat2_results_transposed } + // decompress main bins (and large unbinned contigs) for downstream, + // first have to separate and re-group due to limitation of GUNZIP + METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } + SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } - GUNZIP ( ch_metabat2_results_transposed ) + ch_metabat2_results_transposed.mix( ch_split_fasta_results_transposed ).set { ch_final_bins_for_gunzip } + GUNZIP ( ch_final_bins_for_gunzip ) GUNZIP.out.gunzip.groupTuple(by: 0).set{ ch_metabat_results_gunzipped } // Compute bin depths for different samples (according to `binning_map_mode`) ch_depth_input = ch_metabat_results_gunzipped - .dump(tag: "depth_input") .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) MAG_DEPTHS ( ch_depth_input ) From 70cd3d45483c0e2b51307b4fdd074c7eeda0fcdf Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 24 Jan 2022 17:55:26 +0100 Subject: [PATCH 037/246] fix: updates following code review --- conf/modules.config | 23 ++++++++-- docs/output.md | 8 ++-- modules/local/metabat2.nf | 56 ----------------------- modules/local/split_fasta.nf | 10 ++-- subworkflows/local/binning_preparation.nf | 2 - 5 files changed, 28 insertions(+), 71 deletions(-) delete mode 100644 modules/local/metabat2.nf diff --git a/conf/modules.config b/conf/modules.config index 10cc734a..1bf2f24f 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -198,12 +198,12 @@ process { pattern: 'bins/*.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded_contigs" }, + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded" }, mode: 'copy', pattern: '*tooShort.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded_contigs" }, + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded" }, mode: 'copy', pattern: '*lowDepth.fa.gz' ] @@ -218,8 +218,23 @@ process { withName: SPLIT_FASTA { publishDir = [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/" }, - mode: 'copy', + publishDir = [ + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/unbinned" }, + mode: 'copy', + pattern: '*unbinned.[0-9]*.fa.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded/" }, + mode: 'copy', + pattern: '*unbinned.pooled.fa.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded/" }, + mode: 'copy', + pattern: '*unbinned.remaining.fa.gz' + ] + ] ] } diff --git a/docs/output.md b/docs/output.md index 6a6275e7..ad2ad311 100644 --- a/docs/output.md +++ b/docs/output.md @@ -241,7 +241,7 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig

Output files * `GenomeBinning/` - * `[sample/group]-[assembler]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads. + * `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads. @@ -253,8 +253,9 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig Output files * `GenomeBinning/MetaBAT2/` - * `bins/[assembler]-[sample/group].*.fa`: Genome bins retrieved from input assembly - * `discarded/[assembler]-[sample/group].{unbinned,lowDepth,tooShort}.*.fa`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported + * `bins/[assembler]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + * `unbinned/[assembler]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported + * `discarded/[assembler]-[sample/group].{lowDepth,tooShort,unbinned,poooled,unbinned.remaining}.*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. These consist of the ones that did not pass the length filtering thresholds @@ -266,6 +267,7 @@ All the files and contigs in this folder will be assessed by QUAST and BUSCO. * `GenomeBinning/MetaBAT2/discarded/` * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBat2 * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBat2 +* `GenomeBinning/MetaBAT2/unbinned/` * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. diff --git a/modules/local/metabat2.nf b/modules/local/metabat2.nf deleted file mode 100644 index fe017ce9..00000000 --- a/modules/local/metabat2.nf +++ /dev/null @@ -1,56 +0,0 @@ -// Import generic module functions -include { initOptions; saveFiles; getSoftwareName } from './functions' - -params.options = [:] -options = initOptions(params.options) - -process METABAT2 { - tag "${meta.assembler}-${meta.id}" - - publishDir "${params.outdir}", - mode: params.publish_dir_mode, - saveAs: { filename -> saveFiles(filename:filename, options:params.options, publish_dir:getSoftwareName(task.process), meta:meta, publish_by_meta:['assembler']) } - - conda (params.enable_conda ? "bioconda::metabat2=2.15 conda-forge::python=3.6.7 conda-forge::biopython=1.74 conda-forge::pandas=1.1.5" : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container "https://depot.galaxyproject.org/singularity/mulled-v2-e25d1fa2bb6cbacd47a4f8b2308bd01ba38c5dd7:75310f02364a762e6ba5206fcd11d7529534ed6e-0" - } else { - container "quay.io/biocontainers/mulled-v2-e25d1fa2bb6cbacd47a4f8b2308bd01ba38c5dd7:75310f02364a762e6ba5206fcd11d7529534ed6e-0" - } - - input: - tuple val(meta), path(assembly), path(bam), path(bai), path(depth) - - output: - tuple val(meta), path("MetaBAT2/*.fa") , emit: bins - path "${meta.assembler}-${meta.id}-depth.txt.gz" , emit: depths - path "MetaBAT2/discarded/*" , emit: discarded - path '*.version.txt' , emit: version - - script: - def software = getSoftwareName(task.process) - """ - ## OMP_NUM_THREADS=${task.cpus} jgi_summarize_bam_contig_depths --outputDepth depth.txt ${bam} - gunzip -f $depth - - metabat2 -t "${task.cpus}" -i "${assembly}" -a ${depth.getName().replace(".gz", "")} -o "MetaBAT2/${meta.assembler}-${meta.id}" -m ${params.min_contig_size} --unbinned --seed ${params.metabat_rng_seed} - - gzip ${depth.getName().replace(".gz", "")} - mv $depth "${meta.assembler}-${meta.id}-depth.txt.gz" - - # save unbinned contigs above thresholds into individual files, dump others in one file - split_fasta.py "MetaBAT2/${meta.assembler}-${meta.id}.unbinned.fa" ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} - - # delete splitted file so that it doesnt end up in following processes - rm "MetaBAT2/${meta.assembler}-${meta.id}.unbinned.fa" - - mkdir MetaBAT2/discarded - gzip "MetaBAT2/${meta.assembler}-${meta.id}.lowDepth.fa" \ - "MetaBAT2/${meta.assembler}-${meta.id}.tooShort.fa" \ - "MetaBAT2/${meta.assembler}-${meta.id}.unbinned.pooled.fa" \ - "MetaBAT2/${meta.assembler}-${meta.id}.unbinned.remaining.fa" - mv "MetaBAT2/${meta.assembler}-${meta.id}".*.fa.gz MetaBAT2/discarded/ - - echo \$(metabat2 --help 2>&1) | sed "s/^.*version 2\\://; s/ (Bioconda.*//" > ${software}.version.txt - """ -} diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 93ba6431..24465e1f 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -14,17 +14,15 @@ process SPLIT_FASTA { tuple val(meta), path(unbinned) output: - tuple val(meta), path("unbinned/*unbinned.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do - tuple val(meta), path("unbinned/*unbinned.pooled.fa.gz") , optional:true, emit: pooled - tuple val(meta), path("unbinned/*unbinned.remaining.fa.gz") , optional:true, emit: remaining + tuple val(meta), path("*unbinned.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do + tuple val(meta), path("*unbinned.pooled.fa.gz") , optional:true, emit: pooled + tuple val(meta), path("*unbinned.remaining.fa.gz") , optional:true, emit: remaining script: """ # save unbinned contigs above thresholds into individual files, dump others in one file split_fasta.py $unbinned ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} - mkdir -p unbinned/ - mv *.fa unbinned/ - gzip unbinned/* + gzip *.unbinned.*.fa """ } diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf index 96dbf833..d8a8899d 100644 --- a/subworkflows/local/binning_preparation.nf +++ b/subworkflows/local/binning_preparation.nf @@ -44,8 +44,6 @@ workflow BINNING_PREPARATION { .groupTuple(by: 0) .map { meta, assembly, bams, bais -> [ meta, assembly[0], bams, bais ] } // multiple symlinks to the same assembly -> use first - ch_grouped_mappings - emit: bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } bowtie2_version = BOWTIE2_ASSEMBLY_ALIGN.out.version From bf0e3e5683d1ea7b3b392abc2ef86a6f4e5732dd Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Tue, 25 Jan 2022 13:18:15 +0100 Subject: [PATCH 038/246] Apply suggestions from code review Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- docs/output.md | 2 -- 1 file changed, 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index ad2ad311..75f412ac 100644 --- a/docs/output.md +++ b/docs/output.md @@ -255,7 +255,6 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig * `GenomeBinning/MetaBAT2/` * `bins/[assembler]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly * `unbinned/[assembler]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported - * `discarded/[assembler]-[sample/group].{lowDepth,tooShort,unbinned,poooled,unbinned.remaining}.*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. These consist of the ones that did not pass the length filtering thresholds @@ -267,7 +266,6 @@ All the files and contigs in this folder will be assessed by QUAST and BUSCO. * `GenomeBinning/MetaBAT2/discarded/` * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBat2 * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBat2 -* `GenomeBinning/MetaBAT2/unbinned/` * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. From 38dce841a6dc5131644555c60d0a1f3be20f22f0 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 25 Jan 2022 13:26:27 +0100 Subject: [PATCH 039/246] fix: update from code review --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 75f412ac..40236bc4 100644 --- a/docs/output.md +++ b/docs/output.md @@ -258,7 +258,7 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig -All the files and contigs in this folder will be assessed by QUAST and BUSCO. +All the files and contigs in these folders will be assessed by QUAST and BUSCO.
Output files From ddcd7d944d2731dbfbae74201fcca714d253b599 Mon Sep 17 00:00:00 2001 From: maxibor Date: Thu, 27 Jan 2022 21:52:34 +0100 Subject: [PATCH 040/246] fix linting --- CHANGELOG.md | 1 + nextflow.config | 6 +++--- nextflow_schema.json | 2 +- subworkflows/local/ancient_dna.nf | 28 ++++++++++++++-------------- workflows/mag.nf | 10 +++++----- 5 files changed, 24 insertions(+), 23 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 96198e5a..c6ac24d0 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -6,6 +6,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ## v2.2.0dev ### `Added` + - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow diff --git a/nextflow.config b/nextflow.config index 37aef95c..05f042df 100644 --- a/nextflow.config +++ b/nextflow.config @@ -26,7 +26,7 @@ params { host_removal_save_ids = false // binning options - bowtie2_mode = '' + bowtie2_mode = null binning_map_mode = 'group' skip_binning = false min_contig_size = 1500 @@ -36,8 +36,8 @@ params { // assembly options coassemble_group = false - spades_options = '' - megahit_options = '' + spades_options = null + megahit_options = null skip_spades = false skip_spadeshybrid = false skip_megahit = false diff --git a/nextflow_schema.json b/nextflow_schema.json index 99301202..676aa9e5 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -551,7 +551,7 @@ "bowtie2_mode": { "type": "string", "description": "Bowtie2 alignment mode", - "help_text": "Bowtie2 alignment mode options, for example: `--very fast` , `--very-sensitive -N 1` , ..." + "help_text": "Bowtie2 alignment mode options, for example: `--very-fast` , `--very-sensitive-local -N 1` , ..." }, "skip_prokka": { "type": "boolean", diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index 0b191b7f..e502c35a 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -1,10 +1,10 @@ -include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main' -include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main' -include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main' -include { FREEBAYES } from '../../modules/nf-core/modules/freebayes/main' -include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main' -include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main' -include { SAMTOOLS_FAIDX as FAIDX} from '../../modules/nf-core/modules/samtools/faidx/main' +include { BCFTOOLS_CONSENSUS } from '../../modules/nf-core/modules/bcftools/consensus/main' +include { BCFTOOLS_INDEX as BCFTOOLS_INDEX_PRE ; BCFTOOLS_INDEX as BCFTOOLS_INDEX_POST } from '../../modules/nf-core/modules/bcftools/index/main' +include { BCFTOOLS_VIEW } from '../../modules/nf-core/modules/bcftools/view/main' +include { FREEBAYES } from '../../modules/nf-core/modules/freebayes/main' +include { PYDAMAGE_ANALYZE } from '../../modules/nf-core/modules/pydamage/analyze/main' +include { PYDAMAGE_FILTER } from '../../modules/nf-core/modules/pydamage/filter/main' +include { SAMTOOLS_FAIDX as FAIDX} from '../../modules/nf-core/modules/samtools/faidx/main' workflow ANCIENT_DNA_ASSEMLY_VALIDATION { take: @@ -15,13 +15,13 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { FAIDX(input.map { item -> [ item[0], item[1] ] }) input.join(FAIDX.out.fai) .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] - FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], []] }, - freebayes_input.map { item -> item[1] }, - freebayes_input.map { item -> item[4] }, - [], - [], - [], - [] ) + FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], []] }, + freebayes_input.map { item -> item[1] }, + freebayes_input.map { item -> item[4] }, + [], + [], + [], + [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) BCFTOOLS_VIEW(FREEBAYES.out.vcf.join(BCFTOOLS_INDEX_PRE.out.tbi), [], [], []) diff --git a/workflows/mag.nf b/workflows/mag.nf index 6a07680e..f9fa7cd0 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -488,16 +488,16 @@ workflow MAG { /* ================================================================================ - Binning preparation + Binning preparation ================================================================================ */ - + ch_bowtie2_assembly_multiqc = Channel.empty() ch_busco_summary = Channel.empty() ch_busco_multiqc = Channel.empty() - + BINNING_PREPARATION ( ch_assemblies, @@ -522,7 +522,7 @@ workflow MAG { */ if (!params.skip_binning){ - + if (params.ancient_dna) { METABAT2_BINNING ( BINNING_PREPARATION.out.grouped_mappings @@ -537,7 +537,7 @@ workflow MAG { ) } - + ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_software_versions = ch_software_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first().ifEmpty(null)) ch_software_versions = ch_software_versions.mix(METABAT2_BINNING.out.metabat2_version.first().ifEmpty(null)) From 467e2699791b90fa99587d3379ed6491d103537a Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 09:55:44 +0100 Subject: [PATCH 041/246] back to non-standard default for null for now --- nextflow.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index 05f042df..d92a0f43 100644 --- a/nextflow.config +++ b/nextflow.config @@ -36,8 +36,8 @@ params { // assembly options coassemble_group = false - spades_options = null - megahit_options = null + spades_options = '' + megahit_options = '' skip_spades = false skip_spadeshybrid = false skip_megahit = false From 1edb694efa26df48d66bda8ebeceea49b184a326 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 12:00:46 +0100 Subject: [PATCH 042/246] remove aDNA profile mention --- docs/usage.md | 6 ------ 1 file changed, 6 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 4af31795..a89959c8 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -66,10 +66,6 @@ Please note the following requirements: Again, by default, the group information is only used to compute co-abundances for the binning step, but not for group-wise co-assembly (see the parameter docs for [`--coassemble_group`](https://nf-co.re/mag/parameters#coassemble_group) and [`--binning_map_mode`](https://nf-co.re/mag/parameters#binning_map_mode) for more information about how this group information can be used). -### Ancient DNA mode - -Using the ancient DNA profile `-profile ancient_dna` (see [`profile` section](https://nf-co.re/mag/2.1.0/usage#profile)), you can perform ancient DNA *de novo* assembly validation. - ## Running the pipeline The typical command for running the pipeline is as follows: @@ -152,8 +148,6 @@ If `-profile` is not specified, the pipeline will run locally and expect all sof * `test`, `test_hybrid`, `test_host_rm`, `test_hybrid_host_rm`, `test_busco_auto` * Profiles with a complete configuration for automated testing * Includes links to test data so needs no other parameters -* `ancient_dna` - * A profile to run nf-core/mag with ancient DNA assembly specific parameters and the ancient DNA subworkflow. ### `-resume` From dce284eca03560f23c4f4ad590ac71dcef3be7f8 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 13:25:14 +0100 Subject: [PATCH 043/246] add aDNA workflow description --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 676aa9e5..1548cbae 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -593,7 +593,7 @@ "ancient_dna_assembly": { "title": "Ancient DNA assembly", "type": "object", - "description": "", + "description": "Performs ancient DNA assembly validation and contig consensus sequence recalling.", "default": "", "properties": { "ancient_dna": { From 6e3a92e576c56d17d2d613162a9ae814b790df50 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 14:48:37 +0100 Subject: [PATCH 044/246] remove useless header --- docs/usage.md | 2 -- 1 file changed, 2 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index a89959c8..0b7c436a 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -159,8 +159,6 @@ You can also supply a run name to resume a specific run: `-resume [run-name]`. U Specify the path to a specific config file (this is a core Nextflow command). See the [nf-core website documentation](https://nf-co.re/usage/configuration) for more information. -## Custom configuration - ### Resource requests Whilst the default requirements set within the pipeline will hopefully work for most people and with most input data, you may find that you want to customise the compute resources that the pipeline requests. Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with any of the error codes specified [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L18) it will automatically be resubmitted with higher requests (2 x original, then 3 x original). If it still fails after the third attempt then the pipeline execution is stopped. From 6b3258c88fedf9bbe5162dd723912063fdf2ef87 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 16:14:15 +0100 Subject: [PATCH 045/246] update freebayes modules --- conf/test_ancient_dna.config | 2 +- modules.json | 2 +- modules/nf-core/modules/freebayes/main.nf | 6 +++--- 3 files changed, 5 insertions(+), 5 deletions(-) diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config index 2f852e36..550dcb7f 100644 --- a/conf/test_ancient_dna.config +++ b/conf/test_ancient_dna.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test, + nextflow run nf-core/mag -profile test_ancient_dna, ---------------------------------------------------------------------------------------- */ diff --git a/modules.json b/modules.json index 7d954554..7f0a05b0 100644 --- a/modules.json +++ b/modules.json @@ -19,7 +19,7 @@ "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, "freebayes": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" }, "gunzip": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" diff --git a/modules/nf-core/modules/freebayes/main.nf b/modules/nf-core/modules/freebayes/main.nf index 0568ca88..1dd91fef 100644 --- a/modules/nf-core/modules/freebayes/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -22,7 +22,7 @@ process FREEBAYES { script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" def input = input_2 ? "${input_1} ${input_2}" : "${input_1}" def targets_file = targets ? "--target ${targets}" : "" def samples_file = samples ? "--samples ${samples}" : "" @@ -41,7 +41,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - bgzip ${prefix}.vcf + gzip --no-name ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": @@ -60,7 +60,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - bgzip ${prefix}.vcf + gzip --no-name ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": From 81291721d4b82795d50a317045b55fb1d97ee3a3 Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 16:24:50 +0100 Subject: [PATCH 046/246] add aDNA to README --- README.md | 1 + 1 file changed, 1 insertion(+) diff --git a/README.md b/README.md index 895161ed..8dcf8409 100644 --- a/README.md +++ b/README.md @@ -37,6 +37,7 @@ The pipeline then: * predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) * performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) * assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) +* performs (optionaly) ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html). Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. 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a/docs/images/mag_workflow.svg b/docs/images/mag_workflow.svg index 7f78e0fc..2d003098 100644 --- a/docs/images/mag_workflow.svg +++ b/docs/images/mag_workflow.svg @@ -5,9 +5,9 @@ viewBox="0 0 310.4236 140.53292" version="1.1" id="svg8" - inkscape:version="1.1.1 (c3084ef, 2021-09-22)" - sodipodi:docname="mag_workflow.svg" - inkscape:export-filename="mag_workflow.png" + inkscape:version="1.1.1 (1:1.1+202109281949+c3084ef5ed)" + sodipodi:docname="mag_workflow_new.svg" + inkscape:export-filename="mag_workflow_new.png" inkscape:export-xdpi="289.40701" inkscape:export-ydpi="289.40701" xmlns:inkscape="http://www.inkscape.org/namespaces/inkscape" @@ -319,7 +319,7 @@ y2="103.70081" gradientUnits="userSpaceOnUse" /> Host read removal + style="fill:#24af63;fill-opacity:1;stroke:#000000;stroke-width:0.3;stroke-linecap:square;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> + style="fill:#24af63;fill-opacity:1;stroke:#000000;stroke-width:0.3;stroke-linecap:square;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> Centrifuge - - - Kraken2 - + + Kraken2 + style="fill:#24af63;fill-opacity:1;stroke:#000000;stroke-width:0.3;stroke-linecap:square;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> + style="fill:#24af63;fill-opacity:1;stroke:#000000;stroke-width:0.3;stroke-linecap:square;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> classification (sample- or group-wise) + transform="translate(-127.9581,2.5653297)"> Evaluation QUAST + aDNA Validation + + pyDamage + + Freebayes + + BCFTools @@ -1872,7 +1935,7 @@ ry="2.4730365" /> Date: Fri, 28 Jan 2022 16:45:52 +0100 Subject: [PATCH 048/246] change pipeline summary list ordering --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 8dcf8409..26cdee9a 100644 --- a/README.md +++ b/README.md @@ -34,10 +34,10 @@ The pipeline then: * assigns taxonomy to reads using [Centrifuge](https://ccb.jhu.edu/software/centrifuge/) and/or [Kraken2](https://github.com/DerrickWood/kraken2/wiki) * performs assembly using [MEGAHIT](https://github.com/voutcn/megahit) and [SPAdes](http://cab.spbu.ru/software/spades/), and checks their quality using [Quast](http://quast.sourceforge.net/quast) +* performs (optionaly) ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) * predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) * performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) * assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) -* performs (optionaly) ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html). Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. From ec58993ed1b38c3570c055844ecc8af4e0ef5cad Mon Sep 17 00:00:00 2001 From: maxibor Date: Fri, 28 Jan 2022 17:47:13 +0100 Subject: [PATCH 049/246] move back to bgzip --- modules/nf-core/modules/freebayes/main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/nf-core/modules/freebayes/main.nf b/modules/nf-core/modules/freebayes/main.nf index 1dd91fef..d462088a 100644 --- a/modules/nf-core/modules/freebayes/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -41,7 +41,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - gzip --no-name ${prefix}.vcf + bgzip ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": @@ -60,7 +60,7 @@ process FREEBAYES { $args \\ $input > ${prefix}.vcf - gzip --no-name ${prefix}.vcf + bgzip ${prefix}.vcf cat <<-END_VERSIONS > versions.yml "${task.process}": From b1be7a4a833c2c3d2d003f8e9e1e2f01e39bd847 Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 31 Jan 2022 10:14:18 +0100 Subject: [PATCH 050/246] update freebayes after merged PR --- modules.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules.json b/modules.json index 7f0a05b0..a5806876 100644 --- a/modules.json +++ b/modules.json @@ -19,7 +19,7 @@ "git_sha": "e937c7950af70930d1f34bb961403d9d2aa81c7d" }, "freebayes": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "7026122ec8696016b0b81496d9d39a7918cdb5b2" }, "gunzip": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" From a8c02c9de86ccc4c054633d45295dfd99e7bed2d Mon Sep 17 00:00:00 2001 From: Maxime Borry Date: Mon, 31 Jan 2022 12:30:45 +0100 Subject: [PATCH 051/246] Update README.md Co-authored-by: James A. Fellows Yates --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 26cdee9a..c8fc816f 100644 --- a/README.md +++ b/README.md @@ -34,7 +34,7 @@ The pipeline then: * assigns taxonomy to reads using [Centrifuge](https://ccb.jhu.edu/software/centrifuge/) and/or [Kraken2](https://github.com/DerrickWood/kraken2/wiki) * performs assembly using [MEGAHIT](https://github.com/voutcn/megahit) and [SPAdes](http://cab.spbu.ru/software/spades/), and checks their quality using [Quast](http://quast.sourceforge.net/quast) -* performs (optionaly) ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) +* (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) * predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) * performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) * assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) From e04ba41a795e424e93be32cc879f254b7480fbfd Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 31 Jan 2022 14:19:44 +0100 Subject: [PATCH 052/246] add aDNA subworkflow description in usage docs --- docs/usage.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index 0b7c436a..43821bfa 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -309,3 +309,8 @@ We recommend adding the following line to your environment to limit this (typica ```console NXF_OPTS='-Xms1g -Xmx4g' ``` + +## A note on the ancient DNA subworkflow + +From version 2.2, MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: +[Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can tell appart ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. From f90f3db7315f3cc582f836f21fcef52022642f64 Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 31 Jan 2022 14:22:26 +0100 Subject: [PATCH 053/246] satisfy EClint --- docs/usage.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index 43821bfa..4267e225 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -312,5 +312,6 @@ NXF_OPTS='-Xms1g -Xmx4g' ## A note on the ancient DNA subworkflow -From version 2.2, MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: +From version 2.2, MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: + [Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can tell appart ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. From beeb03cfb02df2e4300df512ad67cbd595e405fe Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 31 Jan 2022 14:28:47 +0100 Subject: [PATCH 054/246] rephasing --- docs/usage.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 4267e225..6801ec4b 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -312,6 +312,6 @@ NXF_OPTS='-Xms1g -Xmx4g' ## A note on the ancient DNA subworkflow -From version 2.2, MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: +MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: -[Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can tell appart ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. +[Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. From 6578ddfa835c253a63b7b34e3b294b4ab975684c Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 2 Feb 2022 11:12:21 +0100 Subject: [PATCH 055/246] fix trailing whitespaces --- conf/modules.config | 6 +++--- nextflow.config | 6 +++--- 2 files changed, 6 insertions(+), 6 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 56f91189..fceb788e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -128,10 +128,10 @@ process { publishDir = [ path: { "${params.outdir}/Assembly" }, mode: 'copy', - saveAs: { - filename -> filename.equals('versions.yml') ? null : + saveAs: { + filename -> filename.equals('versions.yml') ? null : filename.indexOf('.contigs.fa.gz') > 0 ? filename : - filename.indexOf('.log') > 0 ? filename : null } + filename.indexOf('.log') > 0 ? filename : null } ] } diff --git a/nextflow.config b/nextflow.config index 41a3aa16..aec183dd 100644 --- a/nextflow.config +++ b/nextflow.config @@ -36,8 +36,8 @@ params { // assembly options coassemble_group = false - spades_options = '' - megahit_options = '' + spades_options = null + megahit_options = null skip_spades = false skip_spadeshybrid = false skip_megahit = false @@ -199,7 +199,7 @@ if (!params.igenomes_ignore) { } // Export these variables to prevent local Python/R libraries from conflicting with those in the container -// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. +// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. // See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. 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zXwhtT%ei{n#FzEH|C;yZ>+$!u_x#*+`=L8{b9SH^9&27u3G_Gxqxe`L2UJtdxghk z&-wzDFvLvW{chK5u3{n6GSKKy!P&C6w^IFpbD0bcp^A{{2lcLh_DXj@ybtYvc^;(2 M)78&qol`;+0Fu7JivR!s From a3f3d2909577b151e35f7b6b701a32bf11b4bef3 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 2 Feb 2022 11:21:43 +0100 Subject: [PATCH 058/246] fix test profile --- .github/workflows/ci.yml | 5 ----- 1 file changed, 5 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index fddd1a96..45d45ad8 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -52,9 +52,6 @@ jobs: # Only run on push if this is the nf-core dev branch (merged PRs) if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/mag') }} runs-on: ubuntu-latest - env: - NXF_VER: '21.04.0' - NXF_ANSI_LOG: false strategy: matrix: # Run remaining test profiles with minimum nextflow version @@ -64,8 +61,6 @@ jobs: uses: actions/checkout@v2 - name: Install Nextflow - env: - CAPSULE_LOG: none run: | wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ From 0a59dabdb028333a43ce1aec18e9f7784449da07 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 2 Feb 2022 12:36:28 +0100 Subject: [PATCH 059/246] fix speades_options and megahit_options --- modules/local/megahit.nf | 3 ++- modules/local/spades.nf | 3 ++- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/modules/local/megahit.nf b/modules/local/megahit.nf index 6da73ca5..6d9b4b67 100644 --- a/modules/local/megahit.nf +++ b/modules/local/megahit.nf @@ -17,10 +17,11 @@ process MEGAHIT { script: def input = params.single_end ? "-r \"" + reads1.join(",") + "\"" : "-1 \"" + reads1.join(",") + "\" -2 \"" + reads2.join(",") + "\"" + def megahit_options = params.megahit_options ?: '' mem = task.memory.toBytes() if ( !params.megahit_fix_cpu_1 || task.cpus == 1 ) """ - megahit ${params.megahit_options} -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" + megahit $megahit_options -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" gzip -c "MEGAHIT/${meta.id}.contigs.fa" > "MEGAHIT/${meta.id}.contigs.fa.gz" cat <<-END_VERSIONS > versions.yml diff --git a/modules/local/spades.nf b/modules/local/spades.nf index b10b37a4..a374180f 100644 --- a/modules/local/spades.nf +++ b/modules/local/spades.nf @@ -18,11 +18,12 @@ process SPADES { path "versions.yml" , emit: versions script: + def spades_options = params.spades_options ?: '' maxmem = task.memory.toGiga() if ( params.spades_fix_cpus == -1 || task.cpus == params.spades_fix_cpus ) """ metaspades.py \ - ${params.spades_options} \ + $spades_options \ --threads "${task.cpus}" \ --memory $maxmem \ --pe1-1 ${reads[0]} \ From e26d9149e2a37352cc6207e331d40764007e5143 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 2 Feb 2022 12:46:09 +0100 Subject: [PATCH 060/246] fix speades_options for SPADESHYBRID --- modules/local/spadeshybrid.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/modules/local/spadeshybrid.nf b/modules/local/spadeshybrid.nf index 2519a8f7..3566330a 100644 --- a/modules/local/spadeshybrid.nf +++ b/modules/local/spadeshybrid.nf @@ -18,11 +18,12 @@ process SPADESHYBRID { path "versions.yml" , emit: versions script: + def spades_options = params.spades_options ?: '' maxmem = task.memory.toGiga() if ( params.spadeshybrid_fix_cpus == -1 || task.cpus == params.spadeshybrid_fix_cpus ) """ metaspades.py \ - ${params.spades_options} \ + $spades_options \ --threads "${task.cpus}" \ --memory $maxmem \ --pe1-1 ${short_reads[0]} \ From 63b4be58a98ff27f527fabb1c5718374840d9870 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 2 Feb 2022 13:34:15 +0100 Subject: [PATCH 061/246] fix containers --- modules/local/bowtie2_assembly_build.nf | 8 +++----- modules/local/bowtie2_removal_build.nf | 8 +++----- 2 files changed, 6 insertions(+), 10 deletions(-) diff --git a/modules/local/bowtie2_assembly_build.nf b/modules/local/bowtie2_assembly_build.nf index 830d68bd..892477c6 100644 --- a/modules/local/bowtie2_assembly_build.nf +++ b/modules/local/bowtie2_assembly_build.nf @@ -2,11 +2,9 @@ process BOWTIE2_ASSEMBLY_BUILD { tag "${meta.assembler}-${meta.id}" conda (params.enable_conda ? 'bioconda::bowtie2=2.4.2' : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container 'https://depot.galaxyproject.org/singularity/bowtie2:2.4.2--py38h1c8e9b9_1' - } else { - container 'quay.io/biocontainers/bowtie2:2.4.2--py38h1c8e9b9_1' - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bowtie2:2.4.2--py38h1c8e9b9_1' : + 'quay.io/biocontainers/bowtie2:2.4.2--py38h1c8e9b9_1' }" input: tuple val(meta), path(assembly) diff --git a/modules/local/bowtie2_removal_build.nf b/modules/local/bowtie2_removal_build.nf index 824eaac2..9f8531dd 100644 --- a/modules/local/bowtie2_removal_build.nf +++ b/modules/local/bowtie2_removal_build.nf @@ -2,11 +2,9 @@ process BOWTIE2_REMOVAL_BUILD { tag "$fasta" conda (params.enable_conda ? 'bioconda::bowtie2=2.4.2' : null) - if (workflow.containerEngine == 'singularity' && !params.singularity_pull_docker_container) { - container 'https://depot.galaxyproject.org/singularity/bowtie2:2.4.2--py38h1c8e9b9_1' - } else { - container 'quay.io/biocontainers/bowtie2:2.4.2--py38h1c8e9b9_1' - } + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bowtie2:2.4.2--py38h1c8e9b9_1' : + 'quay.io/biocontainers/bowtie2:2.4.2--py38h1c8e9b9_1' }" input: path fasta From 075ce9e438227d60c8619d4db11cbdd10a9fd310 Mon Sep 17 00:00:00 2001 From: d4straub Date: Mon, 7 Feb 2022 16:45:34 +0100 Subject: [PATCH 062/246] fix kronatools version output --- modules/local/krona.nf | 2 +- modules/local/krona_db.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/krona.nf b/modules/local/krona.nf index ce90b151..8eafad18 100644 --- a/modules/local/krona.nf +++ b/modules/local/krona.nf @@ -20,7 +20,7 @@ process KRONA { cat <<-END_VERSIONS > versions.yml "${task.process}": - ktImportTaxonomy: \$(ktImportTaxonomy 2>&1 | sed 's/^.*KronaTools //; s/ - ktImportTaxonomy.*//') + ktImportTaxonomy: \$(ktImportTaxonomy 2>&1 | sed -n '/KronaTools /p' | sed 's/^.*KronaTools //; s/ - ktImportTaxonomy.*//') END_VERSIONS """ } diff --git a/modules/local/krona_db.nf b/modules/local/krona_db.nf index 58f2610f..0c9bf3c8 100644 --- a/modules/local/krona_db.nf +++ b/modules/local/krona_db.nf @@ -15,7 +15,7 @@ process KRONA_DB { cat <<-END_VERSIONS > versions.yml "${task.process}": - ktImportTaxonomy: \$(ktImportTaxonomy 2>&1 | sed 's/^.*KronaTools //; s/ - ktImportTaxonomy.*//') + ktImportTaxonomy: \$(ktImportTaxonomy 2>&1 | sed -n '/KronaTools /p' | sed 's/^.*KronaTools //; s/ - ktImportTaxonomy.*//') END_VERSIONS """ } From 7cad1010602e3b54ccc3a10e0dbe1b05943e827b Mon Sep 17 00:00:00 2001 From: d4straub Date: Mon, 7 Feb 2022 17:03:22 +0100 Subject: [PATCH 063/246] fix github linting by setting node-version to 12 --- .github/workflows/linting.yml | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 3b448773..58abc0ba 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -14,7 +14,7 @@ jobs: - uses: actions/checkout@v2 - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '12' - name: Install markdownlint run: npm install -g markdownlint-cli - name: Run Markdownlint @@ -53,7 +53,7 @@ jobs: - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '12' - name: Install editorconfig-checker run: npm install -g editorconfig-checker @@ -67,7 +67,7 @@ jobs: - uses: actions/checkout@v1 - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '12' - name: Install yaml-lint run: npm install -g yaml-lint - name: Run yaml-lint From 532f09678891a150790b22b74cb5fd0b2741d1ee Mon Sep 17 00:00:00 2001 From: d4straub Date: Mon, 7 Feb 2022 17:07:18 +0100 Subject: [PATCH 064/246] fix linting --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index 979377e6..da75c87f 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -194,7 +194,7 @@ Work dir: Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run` ``` -To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN). We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so based on the search results the file we want is `modules/nf-core/software/star/align/main.nf`. If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9). The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements. The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB. Providing you haven't set any other standard nf-core parameters to __cap__ the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB. The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections. +To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN). We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so based on the search results the file we want is `modules/nf-core/software/star/align/main.nf`. If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9). The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements. The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB. Providing you haven't set any other standard nf-core parameters to **cap** the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB. The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections. ```nextflow process { From d02cd385aec274b083085aa0ae74befc5e541b72 Mon Sep 17 00:00:00 2001 From: d4straub Date: Mon, 7 Feb 2022 17:08:56 +0100 Subject: [PATCH 065/246] fix linting 2 --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index da75c87f..6f653d7c 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -276,6 +276,6 @@ NXF_OPTS='-Xms1g -Xmx4g' ## A note on the ancient DNA subworkflow -MAG integrates an additional subworkflow to validate ancient DNA *de novo* assembly: +MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: [Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. From b103f13a06a9154b32c6eb90d611830463defc8c Mon Sep 17 00:00:00 2001 From: d4straub Date: Mon, 7 Feb 2022 17:18:39 +0100 Subject: [PATCH 066/246] setting node-version to 16 --- .github/workflows/linting.yml | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 58abc0ba..9e785bee 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -14,7 +14,7 @@ jobs: - uses: actions/checkout@v2 - uses: actions/setup-node@v1 with: - node-version: '12' + node-version: '16' - name: Install markdownlint run: npm install -g markdownlint-cli - name: Run Markdownlint @@ -53,7 +53,7 @@ jobs: - uses: actions/setup-node@v1 with: - node-version: '12' + node-version: '16' - name: Install editorconfig-checker run: npm install -g editorconfig-checker @@ -67,7 +67,7 @@ jobs: - uses: actions/checkout@v1 - uses: actions/setup-node@v1 with: - node-version: '12' + node-version: '16' - name: Install yaml-lint run: npm install -g yaml-lint - name: Run yaml-lint From 083ad757b3db991aadb1acbbb719bda684f80402 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 9 Feb 2022 16:28:24 +0100 Subject: [PATCH 067/246] fix config for CENTRIFUGE --- conf/modules.config | 1 - 1 file changed, 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index fceb788e..181f6ad0 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -98,7 +98,6 @@ process { } withName: CENTRIFUGE { - ext.args = '--quiet' publishDir = [ path: { "${params.outdir}/Taxonomy/centrifuge/${meta.id}" }, mode: 'copy', From bcfeb8a7d6f0a8ff4379c057b75dc7a641e85fea Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 9 Feb 2022 16:37:37 +0100 Subject: [PATCH 068/246] change version reporting from sed to cat or tar --- modules/local/busco_db_preparation.nf | 2 +- modules/local/cat_db.nf | 2 +- modules/local/centrifuge_db_preparation.nf | 2 +- modules/local/gtdbtk_db_preparation.nf | 2 +- modules/local/kraken2_db_preparation.nf | 2 +- modules/local/pool_paired_reads.nf | 2 +- modules/local/pool_single_reads.nf | 2 +- 7 files changed, 7 insertions(+), 7 deletions(-) diff --git a/modules/local/busco_db_preparation.nf b/modules/local/busco_db_preparation.nf index 63a036b1..9b8de4e3 100644 --- a/modules/local/busco_db_preparation.nf +++ b/modules/local/busco_db_preparation.nf @@ -21,7 +21,7 @@ process BUSCO_DB_PREPARATION { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + tar: \$(tar --version 2>&1 | sed -n 1p | sed 's/tar (GNU tar) //') END_VERSIONS """ } diff --git a/modules/local/cat_db.nf b/modules/local/cat_db.nf index 7f544654..3642c939 100644 --- a/modules/local/cat_db.nf +++ b/modules/local/cat_db.nf @@ -22,7 +22,7 @@ process CAT_DB { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + tar: \$(tar --version 2>&1 | sed -n 1p | sed 's/tar (GNU tar) //') END_VERSIONS """ } diff --git a/modules/local/centrifuge_db_preparation.nf b/modules/local/centrifuge_db_preparation.nf index 9257a1f6..2f1f7c04 100644 --- a/modules/local/centrifuge_db_preparation.nf +++ b/modules/local/centrifuge_db_preparation.nf @@ -18,7 +18,7 @@ process CENTRIFUGE_DB_PREPARATION { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + tar: \$(tar --version 2>&1 | sed -n 1p | sed 's/tar (GNU tar) //') END_VERSIONS """ } diff --git a/modules/local/gtdbtk_db_preparation.nf b/modules/local/gtdbtk_db_preparation.nf index eb5e0300..f2d79053 100644 --- a/modules/local/gtdbtk_db_preparation.nf +++ b/modules/local/gtdbtk_db_preparation.nf @@ -19,7 +19,7 @@ process GTDBTK_DB_PREPARATION { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + tar: \$(tar --version 2>&1 | sed -n 1p | sed 's/tar (GNU tar) //') END_VERSIONS """ } diff --git a/modules/local/kraken2_db_preparation.nf b/modules/local/kraken2_db_preparation.nf index 859f9592..81c294a8 100644 --- a/modules/local/kraken2_db_preparation.nf +++ b/modules/local/kraken2_db_preparation.nf @@ -21,7 +21,7 @@ process KRAKEN2_DB_PREPARATION { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + tar: \$(tar --version 2>&1 | sed -n 1p | sed 's/tar (GNU tar) //') END_VERSIONS """ } diff --git a/modules/local/pool_paired_reads.nf b/modules/local/pool_paired_reads.nf index a0634096..ac7fcb68 100644 --- a/modules/local/pool_paired_reads.nf +++ b/modules/local/pool_paired_reads.nf @@ -20,7 +20,7 @@ process POOL_PAIRED_READS { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + cat: \$(cat --version 2>&1 | sed -n 1p | sed 's/cat (GNU coreutils) //') END_VERSIONS """ } diff --git a/modules/local/pool_single_reads.nf b/modules/local/pool_single_reads.nf index deda95cc..ac165082 100644 --- a/modules/local/pool_single_reads.nf +++ b/modules/local/pool_single_reads.nf @@ -19,7 +19,7 @@ process POOL_SINGLE_READS { cat <<-END_VERSIONS > versions.yml "${task.process}": - sed: \$(sed --version 2>&1 | sed -n 1p | sed 's/sed (GNU sed) //') + cat: \$(cat --version 2>&1 | sed -n 1p | sed 's/cat (GNU coreutils) //') END_VERSIONS """ } From af444b4cb9404eb983d0b320d5295c6c5d362dda Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 9 Feb 2022 16:46:59 +0100 Subject: [PATCH 069/246] move spades and megahit options to modules.config --- conf/modules.config | 3 +++ modules/local/megahit.nf | 4 ++-- modules/local/spades.nf | 4 ++-- modules/local/spadeshybrid.nf | 4 ++-- 4 files changed, 9 insertions(+), 6 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 181f6ad0..2d00ef14 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -124,6 +124,7 @@ process { //pattern: "*.{fa.gz,log}" //'pattern' didnt work, probably because the output is in a folder, solved with 'saveAs' withName: MEGAHIT { + ext.args = params.megahit_options ?: '' publishDir = [ path: { "${params.outdir}/Assembly" }, mode: 'copy', @@ -135,6 +136,7 @@ process { } withName: SPADES { + ext.args = params.spades_options ?: '' publishDir = [ path: { "${params.outdir}/Assembly/SPAdes" }, mode: 'copy', @@ -143,6 +145,7 @@ process { } withName: SPADESHYBRID { + ext.args = params.spades_options ?: '' publishDir = [ path: { "${params.outdir}/Assembly/SPAdesHybrid" }, mode: 'copy', diff --git a/modules/local/megahit.nf b/modules/local/megahit.nf index 6d9b4b67..3671db73 100644 --- a/modules/local/megahit.nf +++ b/modules/local/megahit.nf @@ -16,12 +16,12 @@ process MEGAHIT { path "versions.yml" , emit: versions script: + def args = task.ext.args ?: '' def input = params.single_end ? "-r \"" + reads1.join(",") + "\"" : "-1 \"" + reads1.join(",") + "\" -2 \"" + reads2.join(",") + "\"" - def megahit_options = params.megahit_options ?: '' mem = task.memory.toBytes() if ( !params.megahit_fix_cpu_1 || task.cpus == 1 ) """ - megahit $megahit_options -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" + megahit $args -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" gzip -c "MEGAHIT/${meta.id}.contigs.fa" > "MEGAHIT/${meta.id}.contigs.fa.gz" cat <<-END_VERSIONS > versions.yml diff --git a/modules/local/spades.nf b/modules/local/spades.nf index a374180f..baea2876 100644 --- a/modules/local/spades.nf +++ b/modules/local/spades.nf @@ -18,12 +18,12 @@ process SPADES { path "versions.yml" , emit: versions script: - def spades_options = params.spades_options ?: '' + def args = task.ext.args ?: '' maxmem = task.memory.toGiga() if ( params.spades_fix_cpus == -1 || task.cpus == params.spades_fix_cpus ) """ metaspades.py \ - $spades_options \ + $args \ --threads "${task.cpus}" \ --memory $maxmem \ --pe1-1 ${reads[0]} \ diff --git a/modules/local/spadeshybrid.nf b/modules/local/spadeshybrid.nf index 3566330a..caf22d00 100644 --- a/modules/local/spadeshybrid.nf +++ b/modules/local/spadeshybrid.nf @@ -18,12 +18,12 @@ process SPADESHYBRID { path "versions.yml" , emit: versions script: - def spades_options = params.spades_options ?: '' + def args = task.ext.args ?: '' maxmem = task.memory.toGiga() if ( params.spadeshybrid_fix_cpus == -1 || task.cpus == params.spadeshybrid_fix_cpus ) """ metaspades.py \ - $spades_options \ + $args \ --threads "${task.cpus}" \ --memory $maxmem \ --pe1-1 ${short_reads[0]} \ From ac8cc9b16e6743cd2a2fa42c04289e423f4f9314 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 9 Feb 2022 17:00:07 +0100 Subject: [PATCH 070/246] fix CAT and GTDBTK_CLASSIFY output dir --- conf/modules.config | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 2d00ef14..d47912f6 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -221,16 +221,16 @@ process { withName: CAT { publishDir = [ - path: { "${params.outdir}/Taxonomy/CAT/${meta.id}" }, + path: { "${params.outdir}/Taxonomy/CAT/${meta.assembler}" }, mode: 'copy', - pattern: "*.{log,gz}" + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] } withName: GTDBTK_CLASSIFY { ext.args = "--extension fa" publishDir = [ - path: { "${params.outdir}/Taxonomy/GTDB-Tk/${meta.id}" }, + path: { "${params.outdir}/Taxonomy/GTDB-Tk/${meta.assembler}/${meta.id}" }, mode: 'copy', pattern: "*.{log,tasv,tree.gz,fasta,fasta.gz}" ] From 83e1c37c56f7181853d544f56ae4f4224a0b3bf9 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 11 Feb 2022 15:42:41 +0100 Subject: [PATCH 071/246] First attempt - stuck on split FASTQ output and downstream --- conf/modules.config | 37 +++++++++++--- modules.json | 2 +- modules/local/convert_depths.nf | 28 +++++++++++ modules/local/quast.nf | 2 +- modules/local/quast_bins.nf | 2 +- modules/local/split_fasta.nf | 10 ++-- modules/nf-core/modules/maxbin2/main.nf | 3 ++ nextflow.config | 4 ++ .../local/{metabat2_binning.nf => binning.nf} | 50 ++++++++++++++----- workflows/mag.nf | 22 ++++---- 10 files changed, 121 insertions(+), 39 deletions(-) create mode 100644 modules/local/convert_depths.nf rename subworkflows/local/{metabat2_binning.nf => binning.nf} (72%) diff --git a/conf/modules.config b/conf/modules.config index d47912f6..908adfa9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -329,7 +329,7 @@ process { mode: 'copy', pattern: '*-depth.txt.gz' ] - ext.prefix = { "${meta.assembler}-${meta.id}-depth" } + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}-depth" } } withName: METABAT2_METABAT2 { @@ -350,7 +350,7 @@ process { pattern: '*lowDepth.fa.gz' ] ] - ext.prefix = { "${meta.assembler}-${meta.id}" } + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } ext.args = [ "-m ${params.min_contig_size}", "--unbinned", @@ -358,23 +358,44 @@ process { ].join(' ').trim() } + withName: MAXBIN2 { + publishDir = [ + [ + path: { "${params.outdir}/GenomeBinning/MaxBin2/bins/" }, + mode: 'copy', + pattern: '*.*[0-9].fasta.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MaxBin2/discarded" }, + mode: 'copy', + pattern: '*.tooshort.gz' + ], + [ + path: { "${params.outdir}/GenomeBinning/MaxBin2/unbinned" }, + mode: 'copy', + pattern: '*.noclass.gz' + ] + ] + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + } + withName: SPLIT_FASTA { publishDir = [ publishDir = [ [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/unbinned" }, + path: { "${params.outdir}/GenomeBinning/${meta.assembler}/unbinned" }, mode: 'copy', - pattern: '*unbinned.[0-9]*.fa.gz' + pattern: '*.*[0-9].fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded/" }, + path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" }, mode: 'copy', - pattern: '*unbinned.pooled.fa.gz' + pattern: '*.pooled.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/MetaBAT2/discarded/" }, + path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" }, mode: 'copy', - pattern: '*unbinned.remaining.fa.gz' + pattern: '*.remaining.fa.gz' ] ] ] diff --git a/modules.json b/modules.json index 07442973..acf7967a 100644 --- a/modules.json +++ b/modules.json @@ -28,7 +28,7 @@ "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "maxbin2": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "e20e57f90b6787ac9a010a980cf6ea98bd990046" }, "metabat2/jgisummarizebamcontigdepths": { "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" diff --git a/modules/local/convert_depths.nf b/modules/local/convert_depths.nf new file mode 100644 index 00000000..176a9937 --- /dev/null +++ b/modules/local/convert_depths.nf @@ -0,0 +1,28 @@ +process CONVERT_DEPTHS { + tag "${meta.id}" + + conda (params.enable_conda ? "bioconda::bioawk=1.0" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bioawk:1.0--hed695b0_5' : + 'quay.io/biocontainers/bioawk:1.0--hed695b0_5' }" + + input: + tuple val(meta), path(fasta), path(depth) + + output: + // need to add empty val because representing reads as we dont want maxbin to calculate for us. + // theoretically could move this outside the module for clarity but 🤫 + tuple val(meta), path(fasta), val([]), path("*_mb2_depth.txt"), emit: output + + script: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + gunzip $depth + bioawk -t '{{if (NR > 1){{print \$1, \$3}}}}' ${depth.toString() - '.gz'} > ${prefix}_mb2_depth.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bioawk: \$(bioawk --version | cut -f 3 -d ' ' ) + END_VERSIONS + """ +} diff --git a/modules/local/quast.nf b/modules/local/quast.nf index 8a4b3fab..adb4ef28 100644 --- a/modules/local/quast.nf +++ b/modules/local/quast.nf @@ -1,5 +1,5 @@ process QUAST { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" conda (params.enable_conda ? "bioconda::quast=5.0.2" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? diff --git a/modules/local/quast_bins.nf b/modules/local/quast_bins.nf index 22d9478e..ab062609 100644 --- a/modules/local/quast_bins.nf +++ b/modules/local/quast_bins.nf @@ -1,5 +1,5 @@ process QUAST_BINS { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" conda (params.enable_conda ? "bioconda::quast=5.0.2" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 7bb1decd..01648bd6 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -1,5 +1,5 @@ process SPLIT_FASTA { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" label 'process_low' // Using container from metabat2 process, since this will be anyway already downloaded and contains biopython and pandas @@ -12,9 +12,9 @@ process SPLIT_FASTA { tuple val(meta), path(unbinned) output: - tuple val(meta), path("*unbinned.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do - tuple val(meta), path("*unbinned.pooled.fa.gz") , optional:true, emit: pooled - tuple val(meta), path("*unbinned.remaining.fa.gz") , optional:true, emit: remaining + tuple val(meta), path("*.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do + tuple val(meta), path("*.pooled.fa.gz") , optional:true, emit: pooled + tuple val(meta), path("*.remaining.fa.gz") , optional:true, emit: remaining path "versions.yml" , emit: versions script: @@ -22,7 +22,7 @@ process SPLIT_FASTA { # save unbinned contigs above thresholds into individual files, dump others in one file split_fasta.py $unbinned ${params.min_length_unbinned_contigs} ${params.max_unbinned_contigs} ${params.min_contig_size} - gzip *.unbinned.*.fa + gzip *.fa cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/modules/maxbin2/main.nf b/modules/nf-core/modules/maxbin2/main.nf index 4d384391..7b818d00 100644 --- a/modules/nf-core/modules/maxbin2/main.nf +++ b/modules/nf-core/modules/maxbin2/main.nf @@ -21,6 +21,9 @@ process MAXBIN2 { tuple val(meta), path("*_gene.tar.gz"), emit: marker_genes, optional: true path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" diff --git a/nextflow.config b/nextflow.config index aec183dd..d5481c21 100644 --- a/nextflow.config +++ b/nextflow.config @@ -74,6 +74,10 @@ params { // lambda_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Escherichia_virus_Lambda/all_assembly_versions/GCA_000840245.1_ViralProj14204/GCA_000840245.1_ViralProj14204_genomic.fna.gz" lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" + // binning options + skip_metabat2 = false + skip_maxbin2 = false + // Bin QC skip_busco = false busco_reference = null diff --git a/subworkflows/local/metabat2_binning.nf b/subworkflows/local/binning.nf similarity index 72% rename from subworkflows/local/metabat2_binning.nf rename to subworkflows/local/binning.nf index ec239861..3fc1d7f5 100644 --- a/subworkflows/local/metabat2_binning.nf +++ b/subworkflows/local/binning.nf @@ -1,5 +1,5 @@ /* - * Binning with MetaBAT2 + * Binning with MetaBAT2 and MaxBin2 */ params.mag_depths_options = [:] @@ -8,9 +8,11 @@ params.mag_depths_summary_options = [:] include { METABAT2_METABAT2 } from '../../modules/nf-core/modules/metabat2/metabat2/main' include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' +include { MAXBIN2 } from '../../modules/nf-core/modules/maxbin2/main' include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' -include { SPLIT_FASTA } from '../../modules/local/split_fasta' +include { CONVERT_DEPTHS } from '../../modules/local/convert_depths' +include { SPLIT_FASTA } from '../../modules/local/split_fasta' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) @@ -23,7 +25,7 @@ def getColNo(filename) { return lines[0].split('\t').size() } -workflow METABAT2_BINNING { +workflow BINNING { take: assemblies // channel: [ val(meta), path(assembly), path(bams), path(bais) ] reads // channel: [ val(meta), [ reads ] ] @@ -31,21 +33,20 @@ workflow METABAT2_BINNING { main: ch_summarizedepth_input = assemblies - .map { meta, assembly, bams, bais -> [ meta, bams, bais ] } + .map { meta, assembly, bams, bais -> + meta['binner'] = 'MetaBAT2' + [ meta, bams, bais ] + } METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth - .map { it -> - it[0]['binner'] = 'METABAT2' - [ it[0], it[1] ] - } .set { ch_metabat_depths } ch_metabat2_input = assemblies .map { it -> - it[0]['binner'] = 'METABAT2' + it[0]['binner'] = 'MetaBAT2' [ it[0], it[1], it[2], it[3] ] } @@ -54,15 +55,40 @@ workflow METABAT2_BINNING { [ it[0], it[1], it[4]] } - METABAT2_METABAT2 ( ch_metabat2_input ) + // run binning, with convertion of metabat2 depth files to maxbin2 compatible if necessary + + if ( !params.skip_metabat2 ) { + METABAT2_METABAT2 ( ch_metabat2_input ) + } + + if ( !params.skip_maxbin2 ) { + CONVERT_DEPTHS ( ch_metabat2_input ) + .map { it -> + it[0]['binner'] = 'MaxBin2' + + [ it[0], it[1], it[2], it[3] ] + } + .set { ch_maxbin2_input } + + MAXBIN2 ( ch_maxbin2_input ) + } // split FASTQ - METABAT2_METABAT2.out.unbinned - SPLIT_FASTA ( METABAT2_METABAT2.out.unbinned ) + if ( !params.skip_metabat2 & params.skip_maxbin2 ) { + ch_input_splitfasta = METABAT2_METABAT2.out.unbinned + } else if ( params.skip_metabat2 & !params.skip_maxbin2 ) { + ch_input_splitfasta = MAXBIN2.out.unbinned_fasta + } else { + ch_input_splitfasta = METABAT2_METABAT2.out.unbinned.mix(MAXBIN2.out.unbinned_fasta) + } + + SPLIT_FASTA ( ch_input_splitfasta ) // decompress main bins (and large unbinned contigs) for downstream, // first have to separate and re-group due to limitation of GUNZIP METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } + + // TODO Add maxbin2 here SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } ch_metabat2_results_transposed.mix( ch_split_fasta_results_transposed ).set { ch_final_bins_for_gunzip } diff --git a/workflows/mag.nf b/workflows/mag.nf index 250695a5..f19db043 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -88,7 +88,7 @@ include { MULTIQC } from '../modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' -include { METABAT2_BINNING } from '../subworkflows/local/metabat2_binning' +include { BINNING } from '../subworkflows/local/binning' include { BUSCO_QC } from '../subworkflows/local/busco_qc' include { GTDBTK } from '../subworkflows/local/gtdbtk' include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' @@ -523,14 +523,14 @@ workflow MAG { if (!params.skip_binning){ if (params.ancient_dna) { - METABAT2_BINNING ( + BINNING ( BINNING_PREPARATION.out.grouped_mappings .join(ANCIENT_DNA_ASSEMLY_VALIDATION.out.contigs_recalled) .map{ it -> [ it[0], it[4], it[2], it[3] ] }, // [meta, contigs_recalled, bam, bais] ch_short_reads ) } else { - METABAT2_BINNING ( + BINNING ( BINNING_PREPARATION.out.grouped_mappings, ch_short_reads ) @@ -539,8 +539,8 @@ workflow MAG { ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) - ch_versions = ch_versions.mix(METABAT2_BINNING.out.metabat2_version.first()) - ch_versions = ch_versions.mix(METABAT2_BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first()) + ch_versions = ch_versions.mix(BINNING.out.metabat2_version.first()) + ch_versions = ch_versions.mix(BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first()) if (!params.skip_busco){ /* @@ -549,7 +549,7 @@ workflow MAG { BUSCO_QC ( ch_busco_db_file, ch_busco_download_folder, - METABAT2_BINNING.out.bins.transpose() + BINNING.out.bins.transpose() ) ch_busco_summary = BUSCO_QC.out.summary ch_busco_multiqc = BUSCO_QC.out.multiqc @@ -563,7 +563,7 @@ workflow MAG { ch_quast_bins_summary = Channel.empty() if (!params.skip_quast){ - QUAST_BINS ( METABAT2_BINNING.out.bins ) + QUAST_BINS ( BINNING.out.bins ) ch_versions = ch_versions.mix(QUAST_BINS.out.versions.first()) QUAST_BINS_SUMMARY ( QUAST_BINS.out.quast_bin_summaries.collect() ) ch_quast_bins_summary = QUAST_BINS_SUMMARY.out.summary @@ -581,7 +581,7 @@ workflow MAG { ch_cat_db = CAT_DB_GENERATE.out.db } CAT ( - METABAT2_BINNING.out.bins, + BINNING.out.bins, ch_cat_db ) ch_versions = ch_versions.mix(CAT.out.versions.first()) @@ -592,7 +592,7 @@ workflow MAG { ch_gtdbtk_summary = Channel.empty() if ( gtdb ){ GTDBTK ( - METABAT2_BINNING.out.bins, + BINNING.out.bins, ch_busco_summary, ch_gtdb ) @@ -602,7 +602,7 @@ workflow MAG { if (!params.skip_busco || !params.skip_quast || gtdb){ BIN_SUMMARY ( - METABAT2_BINNING.out.depths_summary, + BINNING.out.depths_summary, ch_busco_summary.ifEmpty([]), ch_quast_bins_summary.ifEmpty([]), ch_gtdbtk_summary.ifEmpty([]) @@ -612,7 +612,7 @@ workflow MAG { /* * Prokka: Genome annotation */ - METABAT2_BINNING.out.bins.transpose() + BINNING.out.bins.transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() From 27170fb8907e37927c5abf979f4cecca344a3928 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 11 Feb 2022 17:17:47 +0100 Subject: [PATCH 072/246] More changes --- conf/modules.config | 10 ++++------ modules/local/quast.nf | 2 +- subworkflows/local/binning.nf | 16 +++++++++------- 3 files changed, 14 insertions(+), 14 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 908adfa9..e07b1173 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -329,7 +329,7 @@ process { mode: 'copy', pattern: '*-depth.txt.gz' ] - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}-depth" } + ext.prefix = { "${meta.assembler}-${meta.id}-depth" } } withName: METABAT2_METABAT2 { @@ -381,24 +381,22 @@ process { withName: SPLIT_FASTA { publishDir = [ - publishDir = [ [ - path: { "${params.outdir}/GenomeBinning/${meta.assembler}/unbinned" }, + path: { "${params.outdir}/GenomeBinning/split_fastq/unbinned" }, mode: 'copy', pattern: '*.*[0-9].fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" }, + path: { "${params.outdir}/GenomeBinning/split_fastq/discarded/" }, mode: 'copy', pattern: '*.pooled.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/${meta.assembler}/discarded/" }, + path: { "${params.outdir}/GenomeBinning/split_fastq/discarded/" }, mode: 'copy', pattern: '*.remaining.fa.gz' ] ] - ] } withName: CUSTOM_DUMPSOFTWAREVERSIONS { diff --git a/modules/local/quast.nf b/modules/local/quast.nf index adb4ef28..8a4b3fab 100644 --- a/modules/local/quast.nf +++ b/modules/local/quast.nf @@ -1,5 +1,5 @@ process QUAST { - tag "${meta.assembler}-${meta.binner}-${meta.id}" + tag "${meta.assembler}-${meta.id}" conda (params.enable_conda ? "bioconda::quast=5.0.2" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 3fc1d7f5..a5dbb4d3 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -32,16 +32,15 @@ workflow BINNING { main: + // TODO is scaffolds meant to go into here? These aren't being labelled correctly if so. ch_summarizedepth_input = assemblies .map { meta, assembly, bams, bais -> - meta['binner'] = 'MetaBAT2' [ meta, bams, bais ] } METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth - .set { ch_metabat_depths } ch_metabat2_input = assemblies @@ -56,6 +55,7 @@ workflow BINNING { } // run binning, with convertion of metabat2 depth files to maxbin2 compatible if necessary + // TODO consider replacing if statemetns with the new `when` modules args - requires latest modules though if ( !params.skip_metabat2 ) { METABAT2_METABAT2 ( ch_metabat2_input ) @@ -84,20 +84,22 @@ workflow BINNING { SPLIT_FASTA ( ch_input_splitfasta ) - // decompress main bins (and large unbinned contigs) for downstream, + // decompress main bins (and large unbinned contigs) for MAG Depths, // first have to separate and re-group due to limitation of GUNZIP METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } + MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } - // TODO Add maxbin2 here SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } - ch_metabat2_results_transposed.mix( ch_split_fasta_results_transposed ).set { ch_final_bins_for_gunzip } + ch_metabat2_results_transposed + .mix( ch_maxbin2_results_transposed, ch_split_fasta_results_transposed ) + .set { ch_final_bins_for_gunzip } GUNZIP ( ch_final_bins_for_gunzip ) - GUNZIP.out.gunzip.groupTuple(by: 0).set{ ch_metabat_results_gunzipped } + GUNZIP.out.gunzip.groupTuple(by: 0).set{ ch_binning_results_gunzipped } // Compute bin depths for different samples (according to `binning_map_mode`) - ch_depth_input = ch_metabat_results_gunzipped + ch_depth_input = ch_binning_results_gunzipped .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) MAG_DEPTHS ( ch_depth_input ) From 571f3879c90b793c8a773ea835b97e132c656321 Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?Alex=20H=C3=BCbner?= Date: Wed, 16 Feb 2022 10:26:18 +0100 Subject: [PATCH 073/246] Update modules.config This adds the option to set more stringent quality parameters to `freebayes` and `bcftools view`. In `freebayes`, it sets the possibility to require a minimum base quality considered for genotyping (`params.freebayes_min_basequality`) and the minimal minor allele frequency (`freebayes_minallelefreq`) that is required for freebayes to consider the variant. In `bcftools view`, it adds the option to set two types of minimum genotype qualities. The first one is for high quality variants (`bcftools_view_high_variant_quality`): these are variants that simply pass the minimum genotype quality threshold. The second one is for medium quality variants: these are variants that pass a lower genotype quality threshold (`params.bcftools_view_medium_variant_quality`) but have a minimal number of allele observations to support the alternative genotype (`bcftools_view_minimal_allelesupport`). --- conf/modules.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index d47912f6..b1f3a5ba 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -273,7 +273,7 @@ process { } withName: FREEBAYES { - ext.args = "-p ${params.freebayes_ploidy}" + ext.args = "-p ${params.freebayes_ploidy} -q ${params.freebayes_min_basequality} -F ${params.freebayes_minallelefreq}" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/freebayes" }, pattern: "*.vcf.gz" @@ -281,7 +281,7 @@ process { } withName: BCFTOOLS_VIEW { - ext.args = "-i '%QUAL>=${params.bcftools_view_variant_quality}'" + ext.args = "-v snps,msnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport}'" ext.suffix = ".filtered" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, From 2bfcdc4480b1dd575a6b115bdf3b2ee006bd2d75 Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?Alex=20H=C3=BCbner?= Date: Wed, 16 Feb 2022 10:33:51 +0100 Subject: [PATCH 074/246] Update nextflow.config Add default parameters for the additional filter parameters for `freebayes` and `bcftools view`. --- nextflow.config | 174 +++++++++++++++++++++++++----------------------- 1 file changed, 89 insertions(+), 85 deletions(-) diff --git a/nextflow.config b/nextflow.config index aec183dd..f1aaf031 100644 --- a/nextflow.config +++ b/nextflow.config @@ -10,118 +10,122 @@ params { // Input options - input = null - single_end = false + input = null + single_end = false // short read preprocessing options - save_trimmed_fail = false - fastp_qualified_quality = 15 - fastp_cut_mean_quality = 15 - keep_phix = false - // phix_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Enterobacteria_phage_phiX174_sensu_lato/all_assembly_versions/GCA_002596845.1_ASM259684v1/GCA_002596845.1_ASM259684v1_genomic.fna.gz" - phix_reference = "${baseDir}/assets/data/GCA_002596845.1_ASM259684v1_genomic.fna.gz" - host_fasta = null - host_genome = null - host_removal_verysensitive = false - host_removal_save_ids = false + save_trimmed_fail = false + fastp_qualified_quality = 15 + fastp_cut_mean_quality = 15 + keep_phix = false + // phix_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Enterobacteria_phage_phiX174_sensu_lato/all_assembly_versions/GCA_002596845.1_ASM259684v1/GCA_002596845.1_ASM259684v1_genomic.fna.gz" + phix_reference = "${baseDir}/assets/data/GCA_002596845.1_ASM259684v1_genomic.fna.gz" + host_fasta = null + host_genome = null + host_removal_verysensitive = false + host_removal_save_ids = false // binning options - bowtie2_mode = null - binning_map_mode = 'group' - skip_binning = false - min_contig_size = 1500 - min_length_unbinned_contigs = 1000000 - max_unbinned_contigs = 100 - skip_prokka = false + bowtie2_mode = null + binning_map_mode = 'group' + skip_binning = false + min_contig_size = 1500 + min_length_unbinned_contigs = 1000000 + max_unbinned_contigs = 100 + skip_prokka = false // assembly options - coassemble_group = false - spades_options = null - megahit_options = null - skip_spades = false - skip_spadeshybrid = false - skip_megahit = false - skip_quast = false - skip_prodigal = false + coassemble_group = false + spades_options = null + megahit_options = null + skip_spades = false + skip_spadeshybrid = false + skip_megahit = false + skip_quast = false + skip_prodigal = false // ancient DNA assembly validation options - ancient_dna = false - freebayes_ploidy = 1 - bcftools_view_variant_quality = 10 - pydamage_accuracy = 0.5 + ancient_dna = false + freebayes_ploidy = 1 + freebayes_min_basequality. = 20 + freebayes_minallelefreq. = 0.33 + bcftools_view_high_variant_quality = 30 + bcftools_view_medium_variant_quality = 20 + bcftools_view_minimal_allelesupport = 3 + pydamage_accuracy = 0.5 // taxonomy options - centrifuge_db = null - kraken2_db = null - skip_krona = false - cat_db = null - cat_db_generate = false - save_cat_db = false - gtdb = "https://data.ace.uq.edu.au/public/gtdb/data/releases/release202/202.0/auxillary_files/gtdbtk_r202_data.tar.gz" - gtdbtk_min_completeness = 50.0 - gtdbtk_max_contamination = 10.0 - gtdbtk_min_perc_aa = 10 - gtdbtk_min_af = 0.65 - gtdbtk_pplacer_cpus = 1 - gtdbtk_pplacer_scratch = true + centrifuge_db = null + kraken2_db = null + skip_krona = false + cat_db = null + cat_db_generate = false + save_cat_db = false + gtdb = "https://data.ace.uq.edu.au/public/gtdb/data/releases/release202/202.0/auxillary_files/gtdbtk_r202_data.tar.gz" + gtdbtk_min_completeness = 50.0 + gtdbtk_max_contamination = 10.0 + gtdbtk_min_perc_aa = 10 + gtdbtk_min_af = 0.65 + gtdbtk_pplacer_cpus = 1 + gtdbtk_pplacer_scratch = true // long read preprocessing options - skip_adapter_trimming = false - keep_lambda = false - longreads_min_length = 1000 - longreads_keep_percent = 90 - longreads_length_weight = 10 - // lambda_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Escherichia_virus_Lambda/all_assembly_versions/GCA_000840245.1_ViralProj14204/GCA_000840245.1_ViralProj14204_genomic.fna.gz" - lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" + skip_adapter_trimming = false + keep_lambda = false + longreads_min_length = 1000 + longreads_keep_percent = 90 + longreads_length_weight = 10 + // lambda_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Escherichia_virus_Lambda/all_assembly_versions/GCA_000840245.1_ViralProj14204/GCA_000840245.1_ViralProj14204_genomic.fna.gz" + lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // Bin QC - skip_busco = false - busco_reference = null - busco_download_path = null - busco_auto_lineage_prok = false - save_busco_reference = false + skip_busco = false + busco_reference = null + busco_download_path = null + busco_auto_lineage_prok = false + save_busco_reference = false // Reproducibility options - megahit_fix_cpu_1 = false - spades_fix_cpus = -1 - spadeshybrid_fix_cpus = -1 - metabat_rng_seed = 1 + megahit_fix_cpu_1 = false + spades_fix_cpus = -1 + spadeshybrid_fix_cpus = -1 + metabat_rng_seed = 1 // References - igenomes_base = 's3://ngi-igenomes/igenomes' - igenomes_ignore = false + igenomes_base = 's3://ngi-igenomes/igenomes' + igenomes_ignore = false // MultiQC options - multiqc_config = null - multiqc_title = null - max_multiqc_email_size = '25.MB' + multiqc_config = null + multiqc_title = null + max_multiqc_email_size = '25.MB' // Boilerplate options - outdir = './results' - tracedir = "${params.outdir}/pipeline_info" - email = null - email_on_fail = null - plaintext_email = false - monochrome_logs = false - help = false - validate_params = true - show_hidden_params = false - schema_ignore_params = 'genomes' - enable_conda = false + outdir = './results' + tracedir = "${params.outdir}/pipeline_info" + email = null + email_on_fail = null + plaintext_email = false + monochrome_logs = false + help = false + validate_params = true + show_hidden_params = false + schema_ignore_params = 'genomes' + enable_conda = false // Config options - custom_config_version = 'master' - custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" - config_profile_description = null - config_profile_contact = null - config_profile_url = null - config_profile_name = null + custom_config_version = 'master' + custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" + config_profile_description = null + config_profile_contact = null + config_profile_url = null + config_profile_name = null // Max resource options // Defaults only, expecting to be overwritten - max_memory = '128.GB' - max_cpus = 16 - max_time = '240.h' + max_memory = '128.GB' + max_cpus = 16 + max_time = '240.h' } From 77447c4db9ac20563b8c47ddd10620ede0b68757 Mon Sep 17 00:00:00 2001 From: Alex Huebner Date: Wed, 16 Feb 2022 11:33:28 +0100 Subject: [PATCH 075/246] Fix parameters names to conform with nf-core lint --- nextflow.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index f1aaf031..376f5790 100644 --- a/nextflow.config +++ b/nextflow.config @@ -47,8 +47,8 @@ params { // ancient DNA assembly validation options ancient_dna = false freebayes_ploidy = 1 - freebayes_min_basequality. = 20 - freebayes_minallelefreq. = 0.33 + freebayes_min_basequality = 20 + freebayes_minallelefreq = 0.33 bcftools_view_high_variant_quality = 30 bcftools_view_medium_variant_quality = 20 bcftools_view_minimal_allelesupport = 3 From ea88d79305dc56654c40de9fe9fe55e5395b7130 Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Wed, 16 Feb 2022 13:02:08 +0100 Subject: [PATCH 076/246] Update conf/modules.config --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index b1f3a5ba..2ca02dd5 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -281,7 +281,7 @@ process { } withName: BCFTOOLS_VIEW { - ext.args = "-v snps,msnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport}'" + ext.args = "-v snps,mnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport}'" ext.suffix = ".filtered" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, From a5c82d9ceca2e439b36b891077a7bfc80f3ede5f Mon Sep 17 00:00:00 2001 From: Alex Huebner Date: Wed, 16 Feb 2022 14:32:31 +0100 Subject: [PATCH 077/246] Add newly added parameters to the nextflow_schema.json --- .github/workflows/linting.yml | 6 +++--- nextflow_schema.json | 28 ++++++++++++++++++++++++---- 2 files changed, 27 insertions(+), 7 deletions(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 9e785bee..3b448773 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -14,7 +14,7 @@ jobs: - uses: actions/checkout@v2 - uses: actions/setup-node@v1 with: - node-version: '16' + node-version: '10' - name: Install markdownlint run: npm install -g markdownlint-cli - name: Run Markdownlint @@ -53,7 +53,7 @@ jobs: - uses: actions/setup-node@v1 with: - node-version: '16' + node-version: '10' - name: Install editorconfig-checker run: npm install -g editorconfig-checker @@ -67,7 +67,7 @@ jobs: - uses: actions/checkout@v1 - uses: actions/setup-node@v1 with: - node-version: '16' + node-version: '10' - name: Install yaml-lint run: npm install -g yaml-lint - name: Run yaml-lint diff --git a/nextflow_schema.json b/nextflow_schema.json index fa334729..37030f64 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -576,10 +576,30 @@ "default": 1, "description": "Ploidy for variant calling" }, - "bcftools_view_variant_quality": { + "freebayes_min_basequality": { "type": "integer", - "default": 10, - "description": "Variant calling quality threshold" + "default": 20, + "description": "minimum base quality required for variant calling" + }, + "freebayes_minallelefreq": { + "type": "number", + "default": 0.33, + "description": "minimum minor allele frequency for considering variants" + }, + "bcftools_view_high_variant_quality": { + "type": "integer", + "default": 30, + "description": "minimum genotype quality for considering a variant high quality" + }, + "bcftools_view_medium_variant_quality": { + "type": "integer", + "default": 20, + "description": "minimum genotype quality for considering a variant medium quality" + }, + "bcftools_view_minimal_allelesupport": { + "type": "integer", + "default": 3, + "description": "minimum number of bases supporting the alternative allele" }, "pydamage_accuracy": { "type": "number", @@ -633,4 +653,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} \ No newline at end of file +} From 6a66c51c3497a8c150f85b4f053f2bd54de1a350 Mon Sep 17 00:00:00 2001 From: Alex Huebner Date: Thu, 17 Feb 2022 10:47:41 +0100 Subject: [PATCH 078/246] Close parathensis of bcftools view filtering command --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 2ca02dd5..865d7f52 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -281,7 +281,7 @@ process { } withName: BCFTOOLS_VIEW { - ext.args = "-v snps,mnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport}'" + ext.args = "-v snps,mnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport})'" ext.suffix = ".filtered" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, From d86486258240c64e59caaac1cb5bb625c1a3baf4 Mon Sep 17 00:00:00 2001 From: alexhbnr Date: Tue, 22 Feb 2022 20:21:53 +0100 Subject: [PATCH 079/246] Fix weird lowering of node-version --- .github/workflows/linting.yml | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 3b448773..9e785bee 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -14,7 +14,7 @@ jobs: - uses: actions/checkout@v2 - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '16' - name: Install markdownlint run: npm install -g markdownlint-cli - name: Run Markdownlint @@ -53,7 +53,7 @@ jobs: - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '16' - name: Install editorconfig-checker run: npm install -g editorconfig-checker @@ -67,7 +67,7 @@ jobs: - uses: actions/checkout@v1 - uses: actions/setup-node@v1 with: - node-version: '10' + node-version: '16' - name: Install yaml-lint run: npm install -g yaml-lint - name: Run yaml-lint From 3e167f6cde92b86edc8bb289efd77c9e53842ce0 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 14:31:26 +0100 Subject: [PATCH 080/246] MAG_DEPTHS working only QUAST_BIN_SUMMARY problem --- conf/modules.config | 10 ++--- modules/local/quast_bins.nf | 4 +- subworkflows/local/binning.nf | 81 ++++++++++++++++++++++++----------- workflows/mag.nf | 4 ++ 4 files changed, 66 insertions(+), 33 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index e07b1173..6b983479 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -350,7 +350,7 @@ process { pattern: '*lowDepth.fa.gz' ] ] - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + ext.prefix = { "${meta.assembler}-MetaBAT2-${meta.id}" } ext.args = [ "-m ${params.min_contig_size}", "--unbinned", @@ -376,23 +376,23 @@ process { pattern: '*.noclass.gz' ] ] - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } } withName: SPLIT_FASTA { publishDir = [ [ - path: { "${params.outdir}/GenomeBinning/split_fastq/unbinned" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered" }, mode: 'copy', pattern: '*.*[0-9].fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/split_fastq/discarded/" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered/discarded" }, mode: 'copy', pattern: '*.pooled.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/split_fastq/discarded/" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered/discarded" }, mode: 'copy', pattern: '*.remaining.fa.gz' ] diff --git a/modules/local/quast_bins.nf b/modules/local/quast_bins.nf index ab062609..714f90d9 100644 --- a/modules/local/quast_bins.nf +++ b/modules/local/quast_bins.nf @@ -21,9 +21,9 @@ process QUAST_BINS { for bin in \"\${bins[@]}\"; do metaquast.py --threads "${task.cpus}" --max-ref-number 0 --rna-finding --gene-finding -l "\${bin}" "\${bin}" -o "QUAST/\${bin}" if ! [ -f "QUAST/${meta.assembler}-${meta.id}-quast_summary.tsv" ]; then - cp "QUAST/\${bin}/transposed_report.tsv" "QUAST/${meta.assembler}-${meta.id}-quast_summary.tsv" + cp "QUAST/\${bin}/transposed_report.tsv" "QUAST/${meta.assembler}-${meta.binner}-${meta.id}-quast_summary.tsv" else - tail -n +2 "QUAST/\${bin}/transposed_report.tsv" >> "QUAST/${meta.assembler}-${meta.id}-quast_summary.tsv" + tail -n +2 "QUAST/\${bin}/transposed_report.tsv" >> "QUAST/${meta.assembler}-${meta.binner}-${meta.id}-quast_summary.tsv" fi done diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index a5dbb4d3..e225d2b4 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -32,48 +32,63 @@ workflow BINNING { main: - // TODO is scaffolds meant to go into here? These aren't being labelled correctly if so. + // TODO are scaffolds meant to go into here? These aren't being labelled + // correctly if so... + + // generate coverage depths for each contig ch_summarizedepth_input = assemblies .map { meta, assembly, bams, bais -> - [ meta, bams, bais ] + def meta_new = meta.clone() + [ meta_new, bams, bais ] } METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth + .map { meta, depths -> + def meta_new = meta.clone() + meta_new['binner'] = 'MetaBAT2' + + [ meta_new, depths ] + } .set { ch_metabat_depths } + // combine depths back with assemblies ch_metabat2_input = assemblies - .map { it -> - it[0]['binner'] = 'MetaBAT2' + .map { meta, contigs, reads, indicies -> + def meta_new = meta.clone() + meta_new['binner'] = 'MetaBAT2' - [ it[0], it[1], it[2], it[3] ] + [ meta_new, contigs, reads, indicies ] } .join( ch_metabat_depths, by: 0 ) - .map { it -> - [ it[0], it[1], it[4]] + .map { meta, contigs, reads, indicies, depths -> + [ meta, contigs, depths ] } - // run binning, with convertion of metabat2 depth files to maxbin2 compatible if necessary - // TODO consider replacing if statemetns with the new `when` modules args - requires latest modules though - - if ( !params.skip_metabat2 ) { - METABAT2_METABAT2 ( ch_metabat2_input ) - } - + // conver metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) - .map { it -> - it[0]['binner'] = 'MaxBin2' + CONVERT_DEPTHS.out.output + .map { meta, contigs, reads, depth -> + def meta_new = meta.clone() + meta_new['binner'] = 'MaxBin2' - [ it[0], it[1], it[2], it[3] ] + [ meta_new, contigs, reads, depth ] } .set { ch_maxbin2_input } + } + // run binning + if ( !params.skip_metabat2 ) { + METABAT2_METABAT2 ( ch_metabat2_input ) + } + + if ( !params.skip_maxbin2 ) { MAXBIN2 ( ch_maxbin2_input ) } - // split FASTQ + // split fastq files, depending if ( !params.skip_metabat2 & params.skip_maxbin2 ) { ch_input_splitfasta = METABAT2_METABAT2.out.unbinned } else if ( params.skip_metabat2 & !params.skip_maxbin2 ) { @@ -84,11 +99,11 @@ workflow BINNING { SPLIT_FASTA ( ch_input_splitfasta ) - // decompress main bins (and large unbinned contigs) for MAG Depths, - // first have to separate and re-group due to limitation of GUNZIP + // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for + // MAG Depths, first have to separate and re-group due to limitation of + // GUNZIP module METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } - SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } ch_metabat2_results_transposed @@ -96,11 +111,21 @@ workflow BINNING { .set { ch_final_bins_for_gunzip } GUNZIP ( ch_final_bins_for_gunzip ) - GUNZIP.out.gunzip.groupTuple(by: 0).set{ ch_binning_results_gunzipped } + GUNZIP.out.gunzip + .set{ ch_binning_results_gunzipped } // Compute bin depths for different samples (according to `binning_map_mode`) - ch_depth_input = ch_binning_results_gunzipped - .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + // Have to remove binner meta for grouping to mix back with original depth + // files, as required for MAG_DEPTHS + // Q: where is maxbin2 noclass? All filtered out already? + ch_binning_results_gunzipped + .map { meta, results -> + def meta_new = meta.clone() + [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] + } + .groupTuple (by: 0 ) + .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) @@ -114,11 +139,15 @@ workflow BINNING { .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) - MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) + // Group final binned contigs per sample for final output + ch_binning_results_gunzipped + .groupTuple(by: 0) + .set{ ch_binning_results_final } + emit: - bins = ch_metabat_results_gunzipped + bins = ch_binning_results_final unbinned = SPLIT_FASTA.out.unbinned tooshort = METABAT2_METABAT2.out.tooshort lowdepth = METABAT2_METABAT2.out.lowdepth diff --git a/workflows/mag.nf b/workflows/mag.nf index f19db043..0cdd408b 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -563,7 +563,11 @@ workflow MAG { ch_quast_bins_summary = Channel.empty() if (!params.skip_quast){ + ch_foo = BINNING.out.bins + ch_foo.dump(tag: "input_QUAST_BINS") QUAST_BINS ( BINNING.out.bins ) + ch_bar = QUAST_BINS.out.quast_bin_summaries + ch_bar.dump(tag: "output_QUAST_BINS") ch_versions = ch_versions.mix(QUAST_BINS.out.versions.first()) QUAST_BINS_SUMMARY ( QUAST_BINS.out.quast_bin_summaries.collect() ) ch_quast_bins_summary = QUAST_BINS_SUMMARY.out.summary From 3a1445d0a75210e521d97b886776c665576a6a46 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 14:50:16 +0100 Subject: [PATCH 081/246] Update nextflow schema and remove debugging stuff --- modules/local/convert_depths.nf | 1 - nextflow_schema.json | 10 +++++++++- subworkflows/local/binning.nf | 2 +- workflows/mag.nf | 4 ---- 4 files changed, 10 insertions(+), 7 deletions(-) diff --git a/modules/local/convert_depths.nf b/modules/local/convert_depths.nf index 176a9937..1f5258bf 100644 --- a/modules/local/convert_depths.nf +++ b/modules/local/convert_depths.nf @@ -11,7 +11,6 @@ process CONVERT_DEPTHS { output: // need to add empty val because representing reads as we dont want maxbin to calculate for us. - // theoretically could move this outside the module for clarity but 🤫 tuple val(meta), path(fasta), val([]), path("*_mb2_depth.txt"), emit: output script: diff --git a/nextflow_schema.json b/nextflow_schema.json index fa334729..812ed6d9 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -499,7 +499,15 @@ }, "skip_binning": { "type": "boolean", - "description": "Skip metagenome binning." + "description": "Skip metagenome binning entirely" + }, + "skip_metabat2": { + "type": "boolean", + "description": "Skip MetaBAT2 Binning" + }, + "skip_maxbin2": { + "type": "boolean", + "description": "Skip MaxBin2 Binning" }, "min_contig_size": { "type": "integer", diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index e225d2b4..920f27cb 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -12,7 +12,7 @@ include { MAXBIN2 } from '../../modules/nf-core/mo include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' include { CONVERT_DEPTHS } from '../../modules/local/convert_depths' -include { SPLIT_FASTA } from '../../modules/local/split_fasta' +include { SPLIT_FASTA } from '../../modules/local/split_fasta' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) include { MAG_DEPTHS_SUMMARY } from '../../modules/local/mag_depths_summary' addParams( options: params.mag_depths_summary_options ) diff --git a/workflows/mag.nf b/workflows/mag.nf index 0cdd408b..f19db043 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -563,11 +563,7 @@ workflow MAG { ch_quast_bins_summary = Channel.empty() if (!params.skip_quast){ - ch_foo = BINNING.out.bins - ch_foo.dump(tag: "input_QUAST_BINS") QUAST_BINS ( BINNING.out.bins ) - ch_bar = QUAST_BINS.out.quast_bin_summaries - ch_bar.dump(tag: "output_QUAST_BINS") ch_versions = ch_versions.mix(QUAST_BINS.out.versions.first()) QUAST_BINS_SUMMARY ( QUAST_BINS.out.quast_bin_summaries.collect() ) ch_quast_bins_summary = QUAST_BINS_SUMMARY.out.summary From 2779b5abe9fc2badc8682d021ebb97b02861f68c Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 15:34:23 +0100 Subject: [PATCH 082/246] Fix binning summary --- modules/local/quast_bins.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/quast_bins.nf b/modules/local/quast_bins.nf index 714f90d9..a7e7d1b3 100644 --- a/modules/local/quast_bins.nf +++ b/modules/local/quast_bins.nf @@ -20,7 +20,7 @@ process QUAST_BINS { IFS=', ' read -r -a bins <<< \"\$BINS\" for bin in \"\${bins[@]}\"; do metaquast.py --threads "${task.cpus}" --max-ref-number 0 --rna-finding --gene-finding -l "\${bin}" "\${bin}" -o "QUAST/\${bin}" - if ! [ -f "QUAST/${meta.assembler}-${meta.id}-quast_summary.tsv" ]; then + if ! [ -f "QUAST/${meta.assembler}-${meta.binner}-${meta.id}-quast_summary.tsv" ]; then cp "QUAST/\${bin}/transposed_report.tsv" "QUAST/${meta.assembler}-${meta.binner}-${meta.id}-quast_summary.tsv" else tail -n +2 "QUAST/\${bin}/transposed_report.tsv" >> "QUAST/${meta.assembler}-${meta.binner}-${meta.id}-quast_summary.tsv" From 9beefd332e79c8daad1041f0f5f9880480e72d7b Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 15:54:22 +0100 Subject: [PATCH 083/246] Add error strategy to allow MaxBin fail when no binning can be performed on a FASTA --- conf/modules.config | 3 +++ subworkflows/local/binning.nf | 6 +----- 2 files changed, 4 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 6b983479..d6d55e82 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -377,6 +377,9 @@ process { ] ] ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } + // if no gene found, will crash so allow ignore so rest of pipeline + // completes but without MaxBin2 results + errorStrategy = { task.exitStatus in [ 255 ] ? 'ignore' : 'retry' } } withName: SPLIT_FASTA { diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 920f27cb..87163f13 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -31,10 +31,6 @@ workflow BINNING { reads // channel: [ val(meta), [ reads ] ] main: - - // TODO are scaffolds meant to go into here? These aren't being labelled - // correctly if so... - // generate coverage depths for each contig ch_summarizedepth_input = assemblies .map { meta, assembly, bams, bais -> @@ -100,7 +96,7 @@ workflow BINNING { SPLIT_FASTA ( ch_input_splitfasta ) // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for - // MAG Depths, first have to separate and re-group due to limitation of + // MAG_DEPTHS, first have to separate and re-group due to limitation of // GUNZIP module METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } From ddd3e29ccd7bb213f5b48b73c638012ae0460fcb Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 16:54:54 +0100 Subject: [PATCH 084/246] Update output docs and sync format. Note: noclass suffix removed from maxbin2 unbinned because split_fastq just strips any 2 extensions from end, and maxbin2 unbinned just ends in .noclass.gz (no fasta) --- conf/modules.config | 5 ----- docs/output.md | 39 ++++++++++++++++++++++++++++++++---- modules/local/busco_plot.nf | 8 ++++---- modules/local/split_fasta.nf | 2 +- 4 files changed, 40 insertions(+), 14 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index d6d55e82..2a921713 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -370,11 +370,6 @@ process { mode: 'copy', pattern: '*.tooshort.gz' ], - [ - path: { "${params.outdir}/GenomeBinning/MaxBin2/unbinned" }, - mode: 'copy', - pattern: '*.noclass.gz' - ] ] ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } // if no gene found, will crash so allow ignore so rest of pipeline diff --git a/docs/output.md b/docs/output.md index 12f09260..50eb03ff 100644 --- a/docs/output.md +++ b/docs/output.md @@ -253,8 +253,8 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig

Output files * `GenomeBinning/MetaBAT2/` - * `bins/[assembler]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned/[assembler]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported + * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported
@@ -264,8 +264,39 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO. Output files * `GenomeBinning/MetaBAT2/discarded/` - * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBat2 - * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBat2 + * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBAT2 + * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBAT2 +* `GenomeBinning/MetaBAT2/unbinned_filtered/discarded/` + * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. + * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. + + + +All the files in this folder contain small and/or unbinned contigs that are not further processed. + +Files in these two folders contain all contigs of an assembly. + +### MaxBin2 + +[MaxBin2](https://sourceforge.net/projects/maxbin2/) recovers genome bins (that is, contigs/scaffolds that all belongs to a same organism) from metagenome assemblies. + +
+Output files + +* `GenomeBinning/MaxBin2/` + * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering. + +
+ +All the files and contigs in these folders will be assessed by QUAST and BUSCO. + +
+Output files + +* `GenomeBinning/MaxBin2/discarded/` + * `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 +* `GenomeBinning/MaxBin2/unbinned_filtered/discarded/` * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. diff --git a/modules/local/busco_plot.nf b/modules/local/busco_plot.nf index f4773259..ecda860c 100644 --- a/modules/local/busco_plot.nf +++ b/modules/local/busco_plot.nf @@ -21,17 +21,17 @@ process BUSCO_PLOT { # replace dots in bin names within summary file names by underscores # currently (BUSCO v5.1.0) generate_plot.py does not allow further dots for sum in ${summaries}; do - [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.id}.(.+).txt ]]; + [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt ]]; mode=\${BASH_REMATCH[1]} db_name=\${BASH_REMATCH[2]} - bin="${meta.assembler}-${meta.id}.\${BASH_REMATCH[3]}" + bin="${meta.assembler}-${meta.binner}-${meta.id}.\${BASH_REMATCH[3]}" bin_new="\${bin//./_}" mv \${sum} short_summary.\${mode}.\${db_name}.\${bin_new}.txt done generate_plot.py --working_directory . - mv busco_figure.png "${meta.assembler}-${meta.id}.\${mode}.\${db_name}.busco_figure.png" - mv busco_figure.R "${meta.assembler}-${meta.id}.\${mode}.\${db_name}.busco_figure.R" + mv busco_figure.png "${meta.assembler}-${meta.binner}-${meta.id}.\${mode}.\${db_name}.busco_figure.png" + mv busco_figure.R "${meta.assembler}-${meta.binner}-${meta.id}.\${mode}.\${db_name}.busco_figure.R" fi cat <<-END_VERSIONS > versions.yml diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 01648bd6..358e0408 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -12,7 +12,7 @@ process SPLIT_FASTA { tuple val(meta), path(unbinned) output: - tuple val(meta), path("*.[0-9]*.fa.gz") , optional:true, emit: unbinned //not happy with this glob, but best I can do + tuple val(meta), path("*.[0-9]*.fa.gz") , optional:true, emit: unbinned tuple val(meta), path("*.pooled.fa.gz") , optional:true, emit: pooled tuple val(meta), path("*.remaining.fa.gz") , optional:true, emit: remaining path "versions.yml" , emit: versions From be48baf1abbd94647a407f4e09f55c519ced12be Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Feb 2022 17:00:43 +0100 Subject: [PATCH 085/246] README, Changelog, Citations, Update workflow diagram --- CHANGELOG.md | 2 ++ CITATIONS.md | 3 +++ README.md | 2 +- docs/images/mag_workflow.png | Bin 547427 -> 554690 bytes docs/images/mag_workflow.svg | 37 ++++++++++++++++++++++++++--------- 5 files changed, 34 insertions(+), 10 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index d09f429a..46c0b961 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,6 +9,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow +- [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool + ### `Changed` diff --git a/CITATIONS.md b/CITATIONS.md index 62728297..8135ec5a 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -44,6 +44,9 @@ * [Krona](https://doi.org/10.1186/1471-2105-12-385) > Ondov, B. D., Bergman, N. H., & Phillippy, A. M. (2011). Interactive metagenomic visualization in a Web browser. BMC bioinformatics, 12(1), 1-10. doi: 10.1186/1471-2105-12-385. +* [MaxBin2](https://doi.org/10.1093/bioinformatics/btv638) + > Yu-Wei, W., Simmons, B. A. & Singer, S. W. (2015) MaxBin 2.0: An Automated Binning Algorithm to Recover Genomes from Multiple Metagenomic Datasets. Bioinformatics 32 (4): 605–7. doi: 10.1093/bioinformatics/btv638. + * [MEGAHIT](https://doi.org/10.1016/j.ymeth.2016.02.020) > Li, D., Luo, R., Liu, C. M., Leung, C. M., Ting, H. F., Sadakane, K., ... & Lam, T. W. (2016). MEGAHIT v1. 0: a fast and scalable metagenome assembler driven by advanced methodologies and community practices. Methods, 102, 3-11. doi: 10.1016/j.ymeth.2016.02.020. diff --git a/README.md b/README.md index 79fff6b8..b86fb617 100644 --- a/README.md +++ b/README.md @@ -36,7 +36,7 @@ The pipeline then: * performs assembly using [MEGAHIT](https://github.com/voutcn/megahit) and [SPAdes](http://cab.spbu.ru/software/spades/), and checks their quality using [Quast](http://quast.sourceforge.net/quast) * (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) * predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) -* performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) +* performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/) and/or with [MaxBin2](https://sourceforge.net/projects/maxbin2/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) * assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. diff --git a/docs/images/mag_workflow.png b/docs/images/mag_workflow.png index 7917f6e3ab0c9905a1714e775799425b1606842e..c8291a1b0a301a58bec6cba379527aced229198f 100644 GIT binary patch literal 554690 zcmdSBhdNNH$64A!M)L z^{n^j^LdT;_Ye4;o7amt=XoB_$Kx9J>wev@6Qn3FO?Zal3<`xJl({SU0KWC2P^ZN4 zPQh>Vp3B~bZ>MeVs@tPb%ukX3j?q7U)d0V|bLFRq!$jpZ_2_Hr5iB$%gYxLocrus1 z6NJMVxIH|=J}GALq>OQIOrNHhYM;QU+GUt$7!P$C%yhq7VVD># 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310.4236 140.53292" version="1.1" id="svg8" - inkscape:version="1.1.1 (1:1.1+202109281949+c3084ef5ed)" - sodipodi:docname="mag_workflow_new.svg" + inkscape:version="1.1.2 (1:1.1+202202050950+0a00cf5339)" + sodipodi:docname="mag_workflow.svg" inkscape:export-filename="mag_workflow_new.png" inkscape:export-xdpi="289.40701" inkscape:export-ydpi="289.40701" @@ -376,16 +376,16 @@ borderopacity="1.0" inkscape:pageopacity="0.0" inkscape:pageshadow="2" - inkscape:zoom="1.1988419" - inkscape:cx="735.29296" - inkscape:cy="174.75199" + inkscape:zoom="0.59942095" + inkscape:cx="379.53295" + inkscape:cy="30.863119" inkscape:document-units="mm" - inkscape:current-layer="g390-7" + inkscape:current-layer="g4134" showgrid="true" inkscape:window-width="1920" inkscape:window-height="1016" - inkscape:window-x="0" - inkscape:window-y="27" + inkscape:window-x="1052" + inkscape:window-y="1107" inkscape:window-maximized="1" showguides="false" inkscape:document-rotation="0" @@ -2238,6 +2238,14 @@ width="31.519596" id="rect4770-6-1-7-0" style="fill:#24af63;fill-opacity:1;stroke:#000000;stroke-width:0.296403;stroke-linecap:square;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> + MetaBAT2 + MaxBin2 + transform="translate(110.93518,-163.87148)"> Date: Thu, 24 Feb 2022 17:07:27 +0100 Subject: [PATCH 086/246] Linting --- CHANGELOG.md | 1 - 1 file changed, 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 46c0b961..96f94b7e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,7 +11,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool - ### `Changed` - [#269](https://github.com/nf-core/mag/pull/269) - Update to nf-core 2.2 `TEMPLATE` From 8e32645f2631c50852a16d00abf9c30e4daafd1c Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Mon, 28 Feb 2022 12:23:23 +0100 Subject: [PATCH 087/246] Apply suggestions from code review Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 50eb03ff..31255fa7 100644 --- a/docs/output.md +++ b/docs/output.md @@ -285,7 +285,7 @@ Files in these two folders contain all contigs of an assembly. * `GenomeBinning/MaxBin2/` * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering. + * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering.

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From 2bd66c2904d31b2555d88775af064c370f6637c0 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 28 Feb 2022 12:25:13 +0100 Subject: [PATCH 088/246] Changes from code review --- conf/modules.config | 8 ++++---- docs/output.md | 8 ++++---- 2 files changed, 8 insertions(+), 8 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 2a921713..7789d405 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -374,23 +374,23 @@ process { ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } // if no gene found, will crash so allow ignore so rest of pipeline // completes but without MaxBin2 results - errorStrategy = { task.exitStatus in [ 255 ] ? 'ignore' : 'retry' } + errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } } withName: SPLIT_FASTA { publishDir = [ [ - path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned" }, mode: 'copy', pattern: '*.*[0-9].fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered/discarded" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned/discarded" }, mode: 'copy', pattern: '*.pooled.fa.gz' ], [ - path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned_filtered/discarded" }, + path: { "${params.outdir}/GenomeBinning/${meta.binner}/unbinned/discarded" }, mode: 'copy', pattern: '*.remaining.fa.gz' ] diff --git a/docs/output.md b/docs/output.md index 50eb03ff..469bc50f 100644 --- a/docs/output.md +++ b/docs/output.md @@ -254,7 +254,7 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig * `GenomeBinning/MetaBAT2/` * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported + * `unbinned/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported @@ -266,7 +266,7 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO. * `GenomeBinning/MetaBAT2/discarded/` * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBAT2 * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBAT2 -* `GenomeBinning/MetaBAT2/unbinned_filtered/discarded/` +* `GenomeBinning/MetaBAT2/unbinned/discarded/` * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. @@ -285,7 +285,7 @@ Files in these two folders contain all contigs of an assembly. * `GenomeBinning/MaxBin2/` * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned_filtered/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering. + * `unbinned/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering. @@ -296,7 +296,7 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO. * `GenomeBinning/MaxBin2/discarded/` * `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 -* `GenomeBinning/MaxBin2/unbinned_filtered/discarded/` +* `GenomeBinning/MaxBin2/unbinned/discarded/` * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. From 527cf4540cb1141c876a6796ecfc81398fd03148 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 28 Feb 2022 13:15:12 +0100 Subject: [PATCH 089/246] fix: Add in maxbin2 versions --- subworkflows/local/binning.nf | 1 + workflows/mag.nf | 1 + 2 files changed, 2 insertions(+) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 87163f13..71fcaafc 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -150,4 +150,5 @@ workflow BINNING { depths_summary = MAG_DEPTHS_SUMMARY.out.summary metabat2_version = METABAT2_METABAT2.out.versions metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions + maxbin2_version = MAXBIN2.out.versions } diff --git a/workflows/mag.nf b/workflows/mag.nf index f19db043..2e19e2f1 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -541,6 +541,7 @@ workflow MAG { ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) ch_versions = ch_versions.mix(BINNING.out.metabat2_version.first()) ch_versions = ch_versions.mix(BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first()) + ch_versions = ch_versions.mix(BINNING.out.maxbin2_version.first()) if (!params.skip_busco){ /* From 235cb4c7d9feb7e078b0438bb1c95a7c454d01db Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 28 Feb 2022 13:38:29 +0100 Subject: [PATCH 090/246] Add unbinned to MaxBin2 output for consistency with MetaBat2 --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 7789d405..f3ef973d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -371,7 +371,7 @@ process { pattern: '*.tooshort.gz' ], ] - ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } + ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}.unbinned" } // if no gene found, will crash so allow ignore so rest of pipeline // completes but without MaxBin2 results errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } From 1dbb7bbcfb1a9c2c3d1f46ca8789cba9a9737e1d Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 1 Mar 2022 14:23:02 +0100 Subject: [PATCH 091/246] Start DAS_Tool --- modules.json | 6 ++ .../nf-core/modules/dastool/dastool/main.nf | 67 +++++++++++++ .../nf-core/modules/dastool/dastool/meta.yml | 99 +++++++++++++++++++ .../modules/dastool/scaffolds2bin/main.nf | 40 ++++++++ .../modules/dastool/scaffolds2bin/meta.yml | 58 +++++++++++ subworkflows/local/binning_refinement.nf | 34 +++++++ 6 files changed, 304 insertions(+) create mode 100644 modules/nf-core/modules/dastool/dastool/main.nf create mode 100644 modules/nf-core/modules/dastool/dastool/meta.yml create mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/main.nf create mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/meta.yml create mode 100644 subworkflows/local/binning_refinement.nf diff --git a/modules.json b/modules.json index acf7967a..9bf0d976 100644 --- a/modules.json +++ b/modules.json @@ -15,6 +15,12 @@ "custom/dumpsoftwareversions": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, + "dastool/dastool": { + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + }, + "dastool/scaffolds2bin": { + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + }, "fastp": { "git_sha": "e751e5040af57e1b4e06ed4e0f3efe6de25c1683" }, diff --git a/modules/nf-core/modules/dastool/dastool/main.nf b/modules/nf-core/modules/dastool/dastool/main.nf new file mode 100644 index 00000000..53dfea19 --- /dev/null +++ b/modules/nf-core/modules/dastool/dastool/main.nf @@ -0,0 +1,67 @@ +process DASTOOL_DASTOOL { + tag "$meta.id" + label 'process_medium' + + conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : + 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + + input: + tuple val(meta), path(contigs), path(bins) + path(proteins) + path(db_directory) + val(search_engine) + + output: + tuple val(meta), path("*.log") , emit: log + tuple val(meta), path("*_summary.txt") , emit: summary + tuple val(meta), path("*_DASTool_scaffolds2bin.txt") , emit: scaffolds2bin + tuple val(meta), path("*.eval") , optional: true, emit: eval + tuple val(meta), path("*_DASTool_bins/*.fa") , optional: true, emit: bins + tuple val(meta), path("*.pdf") , optional: true, emit: pdfs + tuple val(meta), path("*.proteins.faa") , optional: true, emit: fasta_proteins + tuple val(meta), path("*.archaea.scg") , optional: true, emit: fasta_archaea_scg + tuple val(meta), path("*.bacteria.scg") , optional: true, emit: fasta_bacteria_scg + tuple val(meta), path("*.seqlength") , optional: true, emit: seqlength + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def bin_list = bins instanceof List ? bins.join(",") : "$bins" + def engine = search_engine ? "--search_engine $search_engine" : "--search_engine diamond" + def db_dir = db_directory ? "--db_directory $db_directory" : "" + def clean_contigs = contigs.toString() - ".gz" + def decompress_contigs = contigs.toString() == clean_contigs ? "" : "gunzip -q -f $contigs" + def decompress_proteins = proteins ? "gunzip -f $proteins" : "" + def clean_proteins = proteins ? proteins.toString() - ".gz" : "" + def proteins_pred = proteins ? "--proteins $clean_proteins" : "" + + if (! search_engine) { + log.info('[DAS_Tool] Default search engine (USEARCH) is proprietary software and not available in bioconda. Using DIAMOND as alternative.') + } + + """ + $decompress_proteins + $decompress_contigs + + DAS_Tool \\ + $args \\ + $proteins_pred \\ + $db_dir \\ + $engine \\ + -t $task.cpus \\ + --bins $bin_list \\ + -c $clean_contigs \\ + -o $prefix + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/dastool/dastool/meta.yml b/modules/nf-core/modules/dastool/dastool/meta.yml new file mode 100644 index 00000000..a77df9bd --- /dev/null +++ b/modules/nf-core/modules/dastool/dastool/meta.yml @@ -0,0 +1,99 @@ +name: dastool_dastool +description: DAS Tool binning step. +keywords: + - binning + - das tool + - table + - de novo + - bins + - contigs + - assembly + - das_tool +tools: + - dastool: + description: | + DAS Tool is an automated method that integrates the results + of a flexible number of binning algorithms to calculate an optimized, non-redundant + set of bins from a single assembly. + + homepage: https://github.com/cmks/DAS_Tool + documentation: https://github.com/cmks/DAS_Tool + tool_dev_url: https://github.com/cmks/DAS_Tool + doi: "10.1038/s41564-018-0171-1" + licence: ["BSD"] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - contigs: + type: file + description: fasta file + pattern: "*.{fa.gz,fas.gz,fasta.gz}" + - bins: + type: file + description: "Scaffolds2bin tabular file generated with dastool/scaffolds2bin" + pattern: "*.scaffolds2bin.tsv" + - proteins: + type: file + description: Predicted proteins in prodigal fasta format (>scaffoldID_geneNo) + pattern: "*.{fa.gz,fas.gz,fasta.gz}" + - db_directory: + type: file + description: (optional) Directory of single copy gene database. + - search_engine: + type: val + description: Engine used for single copy gene identification. USEARCH is not supported due to it being proprietary [blast/diamond] + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - version: + type: file + description: File containing software version + pattern: "versions.yml" + - log: + type: file + description: Log file of the run + pattern: "*.log" + - summary: + type: file + description: Summary of output bins including quality and completeness estimates + pattern: "*summary.txt" + - scaffolds2bin: + type: file + description: Scaffolds to bin file of output bins + pattern: "*.scaffolds2bin.txt" + - eval: + type: file + description: Quality and completeness estimates of input bin sets + pattern: "*.eval" + - pdfs: + type: file + description: Plots showing the amount of high quality bins and score distribution of bins per method + pattern: "*.pdf" + - fasta_proteins: + type: file + description: Output from prodigal if not already supplied + pattern: "*.proteins.faa" + - fasta_archaea_scg: + type: file + description: Results of archaeal single-copy-gene prediction + pattern: "*.archaea.scg" + - fasta_bacteria_scg: + type: file + description: Results of bacterial single-copy-gene prediction + pattern: "*.bacteria.scg" + - seqlength: + type: file + description: Summary of contig lengths + pattern: "*.seqlength" + +authors: + - "@maxibor" + - "@jfy133" diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf b/modules/nf-core/modules/dastool/scaffolds2bin/main.nf new file mode 100644 index 00000000..6e9fcdc2 --- /dev/null +++ b/modules/nf-core/modules/dastool/scaffolds2bin/main.nf @@ -0,0 +1,40 @@ +process DASTOOL_SCAFFOLDS2BIN { + tag "$meta.id" + label 'process_low' + + conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : + 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + + input: + tuple val(meta), path(fasta) + val(extension) + + output: + tuple val(meta), path("*.tsv"), emit: scaffolds2bin + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def file_extension = extension ? extension : "fasta" + + """ + gunzip -f *.${file_extension}.gz + + Fasta_to_Scaffolds2Bin.sh \\ + $args \\ + -i . \\ + -e $file_extension \\ + > ${prefix}.tsv + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml b/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml new file mode 100644 index 00000000..0bf8618d --- /dev/null +++ b/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml @@ -0,0 +1,58 @@ +name: dastool_scaffolds2bin +description: Helper script to convert a set of bins in fasta format to tabular scaffolds2bin format +keywords: + - binning + - das tool + - table + - de novo + - bins + - contigs + - assembly + - das_tool +tools: + - dastool: + description: | + DAS Tool is an automated method that integrates the results + of a flexible number of binning algorithms to calculate an optimized, non-redundant + set of bins from a single assembly. + + homepage: https://github.com/cmks/DAS_Tool + documentation: https://github.com/cmks/DAS_Tool + tool_dev_url: https://github.com/cmks/DAS_Tool + doi: "10.1038/s41564-018-0171-1" + licence: ["BSD"] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Fasta of list of fasta files recommended to be gathered via with .collect() of bins + pattern: "*.{fa,fas,fasta}" + - binner: + type: val + description: Name of the binning software (optional) + - extension: + type: val + description: Fasta file extension (fa | fas | fasta | ...) + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - scaffolds2bin: + type: file + description: tabular scaffolds2bin file for DAS tool input + pattern: "*.scaffolds2bin.tsv" + +authors: + - "@maxibor" diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf new file mode 100644 index 00000000..fd05021e --- /dev/null +++ b/subworkflows/local/binning_refinement.nf @@ -0,0 +1,34 @@ +/* + * Binning with MetaBAT2 and MaxBin2 + */ + +include { DASTOOL_SCAFFOLDS2BIN } from '../../modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_DASTOOL } from '../../modules/dastool/dastool/main.nf' + +workflow BINNING { + take: + contigs // + bins // channel: [ val(meta), path(bins) ] + + main: + + + // Run on each bin file separately + DASTOOL_SCAFFOLDS2BIN ( METABAT2_METABAT2.out.fasta.collect(), "fa") + + // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) + + // Run DAStool + + + + emit: + bins = ch_binning_results_final + unbinned = SPLIT_FASTA.out.unbinned + tooshort = METABAT2_METABAT2.out.tooshort + lowdepth = METABAT2_METABAT2.out.lowdepth + depths_summary = MAG_DEPTHS_SUMMARY.out.summary + metabat2_version = METABAT2_METABAT2.out.versions + metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions + maxbin2_version = MAXBIN2.out.versions +} From 25e7f85d3dcbe12821d2af72f0fa184856010af4 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 1 Mar 2022 14:47:32 +0100 Subject: [PATCH 092/246] Fix split_fasta naming issue for maxbin2 --- bin/split_fasta.py | 7 ++++--- conf/modules.config | 2 +- docs/output.md | 6 +++--- 3 files changed, 8 insertions(+), 7 deletions(-) diff --git a/bin/split_fasta.py b/bin/split_fasta.py index c9eb6886..07d369a6 100755 --- a/bin/split_fasta.py +++ b/bin/split_fasta.py @@ -10,6 +10,7 @@ from Bio.SeqRecord import SeqRecord from Bio.Alphabet import generic_dna import os +import re # Input input_file = argv[1] @@ -19,10 +20,10 @@ # Base name for file output if input_file.endswith('.gz'): - rm_ext=os.path.splitext(input_file)[0] - out_base = (os.path.splitext(rm_ext)[0]) + rm_ext = input_file.replace(".gz", "") + out_base = out_base = re.sub(r'\.fasta$|\.fa$|\.fna$', '', rm_ext) else: - out_base = (os.path.splitext(input_file)[0]) + out_base = re.sub(r'\.fasta$|\.fa$|\.fna$', '', input_file) # Data structures to separate and store sequences df_above_threshold = pd.DataFrame(columns=['id','seq','length']) diff --git a/conf/modules.config b/conf/modules.config index f3ef973d..7789d405 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -371,7 +371,7 @@ process { pattern: '*.tooshort.gz' ], ] - ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}.unbinned" } + ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } // if no gene found, will crash so allow ignore so rest of pipeline // completes but without MaxBin2 results errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } diff --git a/docs/output.md b/docs/output.md index bb292334..4472db52 100644 --- a/docs/output.md +++ b/docs/output.md @@ -285,7 +285,7 @@ Files in these two folders contain all contigs of an assembly. * `GenomeBinning/MaxBin2/` * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. Note that normally these would be output from MaxBin2 with the `.noclass` suffix prior the additional nf-core/mag filtering. + * `unbinned/[assembler]-[binner]-[sample/group].noclass.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. @@ -297,8 +297,8 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO. * `GenomeBinning/MaxBin2/discarded/` * `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 * `GenomeBinning/MaxBin2/unbinned/discarded/` - * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. - * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. + * `*.noclass.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. + * `*.noclass.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. From 790fd1935fc6566584ad51b7f7bf6c1084e282ff Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 8 Mar 2022 13:55:40 +0100 Subject: [PATCH 093/246] Get mostly working other than sending downstream unbinned files? --- conf/modules.config | 11 ++++ modules/local/megahit.nf | 8 +-- modules/local/spades.nf | 24 ++++---- modules/local/spadeshybrid.nf | 24 ++++---- nextflow.config | 7 ++- subworkflows/local/binning.nf | 12 +++- subworkflows/local/binning_refinement.nf | 78 ++++++++++++++++++------ workflows/mag.nf | 27 +++++--- 8 files changed, 133 insertions(+), 58 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 7789d405..2a54f24b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -133,6 +133,7 @@ process { filename.indexOf('.contigs.fa.gz') > 0 ? filename : filename.indexOf('.log') > 0 ? filename : null } ] + ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: SPADES { @@ -142,6 +143,7 @@ process { mode: 'copy', pattern: "*.{fasta.gz,gfa.gz,log}" ] + ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: SPADESHYBRID { @@ -151,6 +153,7 @@ process { mode: 'copy', pattern: "*.{fasta.gz,gfa.gz,log}" ] + ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: QUAST { @@ -397,6 +400,14 @@ process { ] } + withName: DASTOOL_SCAFFOLDS2BIN { + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + } + + withName: DASTOOL_DASTOOL { + ext.prefix = { "${meta.assembler}-${meta.id}" } + } + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/modules/local/megahit.nf b/modules/local/megahit.nf index 3671db73..ee4e1a6c 100644 --- a/modules/local/megahit.nf +++ b/modules/local/megahit.nf @@ -10,9 +10,9 @@ process MEGAHIT { tuple val(meta), path(reads1), path(reads2) output: - tuple val(meta), path("MEGAHIT/${meta.id}.contigs.fa"), emit: assembly + tuple val(meta), path("MEGAHIT/MEGAHIT-${meta.id}.contigs.fa"), emit: assembly path "MEGAHIT/*.log" , emit: log - path "MEGAHIT/${meta.id}.contigs.fa.gz" , emit: assembly_gz + path "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa.gz" , emit: assembly_gz path "versions.yml" , emit: versions script: @@ -21,8 +21,8 @@ process MEGAHIT { mem = task.memory.toBytes() if ( !params.megahit_fix_cpu_1 || task.cpus == 1 ) """ - megahit $args -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" - gzip -c "MEGAHIT/${meta.id}.contigs.fa" > "MEGAHIT/${meta.id}.contigs.fa.gz" + megahit $args -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "MEGAHIT-${meta.id}" + gzip -c "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa" > "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spades.nf b/modules/local/spades.nf index baea2876..9ac93dce 100644 --- a/modules/local/spades.nf +++ b/modules/local/spades.nf @@ -10,11 +10,11 @@ process SPADES { tuple val(meta), path(reads) output: - tuple val(meta), path("${meta.id}_scaffolds.fasta"), emit: assembly - path "${meta.id}.log" , emit: log - path "${meta.id}_contigs.fasta.gz" , emit: contigs_gz - path "${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz - path "${meta.id}_graph.gfa.gz" , emit: graph + tuple val(meta), path("SPAdes-${meta.id}_scaffolds.fasta"), emit: assembly + path "SPAdes-${meta.id}.log" , emit: log + path "SPAdes-${meta.id}_contigs.fasta.gz" , emit: contigs_gz + path "SPAdes-${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz + path "SPAdes-${meta.id}_graph.gfa.gz" , emit: graph path "versions.yml" , emit: versions script: @@ -29,13 +29,13 @@ process SPADES { --pe1-1 ${reads[0]} \ --pe1-2 ${reads[1]} \ -o spades - mv spades/assembly_graph_with_scaffolds.gfa ${meta.id}_graph.gfa - mv spades/scaffolds.fasta ${meta.id}_scaffolds.fasta - mv spades/contigs.fasta ${meta.id}_contigs.fasta - mv spades/spades.log ${meta.id}.log - gzip "${meta.id}_contigs.fasta" - gzip "${meta.id}_graph.gfa" - gzip -c "${meta.id}_scaffolds.fasta" > "${meta.id}_scaffolds.fasta.gz" + mv spades/assembly_graph_with_scaffolds.gfa SPAdes-${meta.id}_graph.gfa + mv spades/scaffolds.fasta SPAdes-${meta.id}_scaffolds.fasta + mv spades/contigs.fasta SPAdes-${meta.id}_contigs.fasta + mv spades/spades.log SPAdes-${meta.id}.log + gzip "SPAdes-${meta.id}_contigs.fasta" + gzip "SPAdes-${meta.id}_graph.gfa" + gzip -c "SPAdes-${meta.id}_scaffolds.fasta" > "SPAdes-${meta.id}_scaffolds.fasta.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spadeshybrid.nf b/modules/local/spadeshybrid.nf index caf22d00..29cf83b4 100644 --- a/modules/local/spadeshybrid.nf +++ b/modules/local/spadeshybrid.nf @@ -10,11 +10,11 @@ process SPADESHYBRID { tuple val(meta), path(long_reads), path(short_reads) output: - tuple val(meta), path("${meta.id}_scaffolds.fasta"), emit: assembly - path "${meta.id}.log" , emit: log - path "${meta.id}_contigs.fasta.gz" , emit: contigs_gz - path "${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz - path "${meta.id}_graph.gfa.gz" , emit: graph + tuple val(meta), path("SPAdesHybrid-${meta.id}_scaffolds.fasta"), emit: assembly + path "SPAdesHybrid-${meta.id}.log" , emit: log + path "SPAdesHybrid-${meta.id}_contigs.fasta.gz" , emit: contigs_gz + path "SPAdesHybrid-${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz + path "SPAdesHybrid-${meta.id}_graph.gfa.gz" , emit: graph path "versions.yml" , emit: versions script: @@ -30,13 +30,13 @@ process SPADESHYBRID { --pe1-2 ${short_reads[1]} \ --nanopore ${long_reads} \ -o spades - mv spades/assembly_graph_with_scaffolds.gfa ${meta.id}_graph.gfa - mv spades/scaffolds.fasta ${meta.id}_scaffolds.fasta - mv spades/contigs.fasta ${meta.id}_contigs.fasta - mv spades/spades.log ${meta.id}.log - gzip "${meta.id}_contigs.fasta" - gzip "${meta.id}_graph.gfa" - gzip -c "${meta.id}_scaffolds.fasta" > "${meta.id}_scaffolds.fasta.gz" + mv spades/assembly_graph_with_scaffolds.gfa SPAdesHybrid-${meta.id}_graph.gfa + mv spades/scaffolds.fasta SPAdesHybrid-${meta.id}_scaffolds.fasta + mv spades/contigs.fasta SPAdesHybrid-${meta.id}_contigs.fasta + mv spades/spades.log SPAdesHybrid-${meta.id}.log + gzip "SPAdesHybrid-${meta.id}_contigs.fasta" + gzip "SPAdesHybrid-${meta.id}_graph.gfa" + gzip -c "SPAdesHybrid-${meta.id}_scaffolds.fasta" > "SPAdesHybrid-${meta.id}_scaffolds.fasta.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/nextflow.config b/nextflow.config index d5481c21..e7dc72ec 100644 --- a/nextflow.config +++ b/nextflow.config @@ -75,8 +75,11 @@ params { lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // binning options - skip_metabat2 = false - skip_maxbin2 = false + skip_metabat2 = false + skip_maxbin2 = false + + // binning refinement + run_dastool = false // Bin QC skip_busco = false diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 71fcaafc..97ed2e86 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -137,14 +137,22 @@ workflow BINNING { MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) + // Final actual bins for bin refinement + METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.fastas) + .groupTuple(by: 0) + .set{ ch_bins_gz } + // Group final binned contigs per sample for final output ch_binning_results_gunzipped .groupTuple(by: 0) .set{ ch_binning_results_final } + + emit: - bins = ch_binning_results_final - unbinned = SPLIT_FASTA.out.unbinned + fastas = ch_binning_results_final // all output from binning + bins_gz = ch_bins_gz // all only binned FASTAs in gzip (no gzip) + unbinned = SPLIT_FASTA.out.unbinned // only (length) filtered unbinned contigs tooshort = METABAT2_METABAT2.out.tooshort lowdepth = METABAT2_METABAT2.out.lowdepth depths_summary = MAG_DEPTHS_SUMMARY.out.summary diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index fd05021e..df070018 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -1,34 +1,76 @@ /* - * Binning with MetaBAT2 and MaxBin2 + * Bin refinement with DASTool */ -include { DASTOOL_SCAFFOLDS2BIN } from '../../modules/dastool/scaffolds2bin/main.nf' -include { DASTOOL_DASTOOL } from '../../modules/dastool/dastool/main.nf' +include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' -workflow BINNING { +workflow BINNING_REFINEMENT { take: - contigs // - bins // channel: [ val(meta), path(bins) ] + contigs // channel: [ val(meta), path(fasta)) ] + bins // channel: [ val(meta), path(binned_fasta_gz) ] main: + ch_versions = Channel.empty() + // run on each bin file separately due to different suffixes + bins + .dump(tag: "input_to_binrefinement") + .branch{ + maxbin2: it[0]['binner'] == 'MaxBin2' + metabat2: it[0]['binner'] == 'MetaBAT2' + } + .set{ ch_for_scaffolds2bin } - // Run on each bin file separately - DASTOOL_SCAFFOLDS2BIN ( METABAT2_METABAT2.out.fasta.collect(), "fa") + // generate bin files + if ( !params.skip_metabat2 & !params.skip_maxbin2 ) { + DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_for_scaffolds2bin.metabat2, "fa") + DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_for_scaffolds2bin.maxbin2, "fasta") + DASTOOL_SCAFFOLDS2BIN_METABAT2.out.scaffolds2bin + .mix(DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.scaffolds2bin) + .set { ch_s2b_for_dastool } + ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions, DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions ) + } else if ( !params.skip_metabat2 & params.skip_maxbin2 ){ + DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_for_scaffolds2bin.metabat2, "fa") + DASTOOL_SCAFFOLDS2BIN_METABAT2.out.scaffolds2bin.set { ch_s2b_for_dastool } + ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions ) + } else if ( params.skip_metabat2 & !params.skip_maxbin2 ){ + DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_for_scaffolds2bin.maxbin2, "fasta") + DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.scaffolds2bin.set { ch_s2b_for_dastool } + ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions ) + } - // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) + // combine per sample + assembler combination + // TODO scaffolds2bin remove binner from meta, group + // TODO contigs remove binner from meta, group + // mergin into input for DASTool + ch_s2b_for_dastool + .dump(tag: "s2b_prior_meta_reduc") + .map { meta, results -> + def meta_new = meta.clone() + [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] + } + .dump(tag: "s2b_prior_merge") + .groupTuple() + .set{ ch_s2b_for_join } + + contigs.dump(tag: "input_contigs_to_s2b_join").join(ch_s2b_for_join, by: 0 ).set{ ch_input_for_dastool } - // Run DAStool + // TODO now picking up noclass/unbinned? + ch_input_for_dastool.dump(tag: "input_to_dastool") + + // Run DAS_Tool + // TODO offer extra parameters + DASTOOL_DASTOOL( ch_input_for_dastool, [], [], [] ) + + + // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) emit: - bins = ch_binning_results_final - unbinned = SPLIT_FASTA.out.unbinned - tooshort = METABAT2_METABAT2.out.tooshort - lowdepth = METABAT2_METABAT2.out.lowdepth - depths_summary = MAG_DEPTHS_SUMMARY.out.summary - metabat2_version = METABAT2_METABAT2.out.versions - metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions - maxbin2_version = MAXBIN2.out.versions + //refined_bins + versions = ch_versions + } diff --git a/workflows/mag.nf b/workflows/mag.nf index 2e19e2f1..20f40925 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -86,12 +86,13 @@ include { MULTIQC } from '../modules // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' -include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' -include { BINNING } from '../subworkflows/local/binning' -include { BUSCO_QC } from '../subworkflows/local/busco_qc' -include { GTDBTK } from '../subworkflows/local/gtdbtk' -include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' +include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' +include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' +include { BINNING } from '../subworkflows/local/binning' +include { BINNING_REFINEMENT } from '../subworkflows/local/binning_refinement' +include { BUSCO_QC } from '../subworkflows/local/busco_qc' +include { GTDBTK } from '../subworkflows/local/gtdbtk' /* ======================================================================================== @@ -536,13 +537,22 @@ workflow MAG { ) } - ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) ch_versions = ch_versions.mix(BINNING.out.metabat2_version.first()) ch_versions = ch_versions.mix(BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first()) ch_versions = ch_versions.mix(BINNING.out.maxbin2_version.first()) + + /* + * DASTool: Binning refinement + */ + + if ( params.run_dastool ) { + BINNING_REFINEMENT ( ch_assemblies, BINNING.out.bins_gz ) + ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) + } + if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly @@ -550,7 +560,7 @@ workflow MAG { BUSCO_QC ( ch_busco_db_file, ch_busco_download_folder, - BINNING.out.bins.transpose() + BINNING.out.fastas.transpose() ) ch_busco_summary = BUSCO_QC.out.summary ch_busco_multiqc = BUSCO_QC.out.multiqc @@ -629,6 +639,7 @@ workflow MAG { ) ch_versions = ch_versions.mix(PROKKA.out.versions.first()) } + } CUSTOM_DUMPSOFTWAREVERSIONS ( From 5752b85baacfd245d51d0ebb56320740373787fb Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 8 Mar 2022 14:38:31 +0100 Subject: [PATCH 094/246] Add a bunch of debugging dumps --- modules/local/megahit.nf | 8 ++++---- modules/local/spades.nf | 24 +++++++++++------------ modules/local/spadeshybrid.nf | 24 +++++++++++------------ subworkflows/local/binning.nf | 12 ++++++++---- subworkflows/local/binning_preparation.nf | 1 + 5 files changed, 37 insertions(+), 32 deletions(-) diff --git a/modules/local/megahit.nf b/modules/local/megahit.nf index 3671db73..ee4e1a6c 100644 --- a/modules/local/megahit.nf +++ b/modules/local/megahit.nf @@ -10,9 +10,9 @@ process MEGAHIT { tuple val(meta), path(reads1), path(reads2) output: - tuple val(meta), path("MEGAHIT/${meta.id}.contigs.fa"), emit: assembly + tuple val(meta), path("MEGAHIT/MEGAHIT-${meta.id}.contigs.fa"), emit: assembly path "MEGAHIT/*.log" , emit: log - path "MEGAHIT/${meta.id}.contigs.fa.gz" , emit: assembly_gz + path "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa.gz" , emit: assembly_gz path "versions.yml" , emit: versions script: @@ -21,8 +21,8 @@ process MEGAHIT { mem = task.memory.toBytes() if ( !params.megahit_fix_cpu_1 || task.cpus == 1 ) """ - megahit $args -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "${meta.id}" - gzip -c "MEGAHIT/${meta.id}.contigs.fa" > "MEGAHIT/${meta.id}.contigs.fa.gz" + megahit $args -t "${task.cpus}" -m $mem $input -o MEGAHIT --out-prefix "MEGAHIT-${meta.id}" + gzip -c "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa" > "MEGAHIT/MEGAHIT-${meta.id}.contigs.fa.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spades.nf b/modules/local/spades.nf index baea2876..9ac93dce 100644 --- a/modules/local/spades.nf +++ b/modules/local/spades.nf @@ -10,11 +10,11 @@ process SPADES { tuple val(meta), path(reads) output: - tuple val(meta), path("${meta.id}_scaffolds.fasta"), emit: assembly - path "${meta.id}.log" , emit: log - path "${meta.id}_contigs.fasta.gz" , emit: contigs_gz - path "${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz - path "${meta.id}_graph.gfa.gz" , emit: graph + tuple val(meta), path("SPAdes-${meta.id}_scaffolds.fasta"), emit: assembly + path "SPAdes-${meta.id}.log" , emit: log + path "SPAdes-${meta.id}_contigs.fasta.gz" , emit: contigs_gz + path "SPAdes-${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz + path "SPAdes-${meta.id}_graph.gfa.gz" , emit: graph path "versions.yml" , emit: versions script: @@ -29,13 +29,13 @@ process SPADES { --pe1-1 ${reads[0]} \ --pe1-2 ${reads[1]} \ -o spades - mv spades/assembly_graph_with_scaffolds.gfa ${meta.id}_graph.gfa - mv spades/scaffolds.fasta ${meta.id}_scaffolds.fasta - mv spades/contigs.fasta ${meta.id}_contigs.fasta - mv spades/spades.log ${meta.id}.log - gzip "${meta.id}_contigs.fasta" - gzip "${meta.id}_graph.gfa" - gzip -c "${meta.id}_scaffolds.fasta" > "${meta.id}_scaffolds.fasta.gz" + mv spades/assembly_graph_with_scaffolds.gfa SPAdes-${meta.id}_graph.gfa + mv spades/scaffolds.fasta SPAdes-${meta.id}_scaffolds.fasta + mv spades/contigs.fasta SPAdes-${meta.id}_contigs.fasta + mv spades/spades.log SPAdes-${meta.id}.log + gzip "SPAdes-${meta.id}_contigs.fasta" + gzip "SPAdes-${meta.id}_graph.gfa" + gzip -c "SPAdes-${meta.id}_scaffolds.fasta" > "SPAdes-${meta.id}_scaffolds.fasta.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spadeshybrid.nf b/modules/local/spadeshybrid.nf index caf22d00..29cf83b4 100644 --- a/modules/local/spadeshybrid.nf +++ b/modules/local/spadeshybrid.nf @@ -10,11 +10,11 @@ process SPADESHYBRID { tuple val(meta), path(long_reads), path(short_reads) output: - tuple val(meta), path("${meta.id}_scaffolds.fasta"), emit: assembly - path "${meta.id}.log" , emit: log - path "${meta.id}_contigs.fasta.gz" , emit: contigs_gz - path "${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz - path "${meta.id}_graph.gfa.gz" , emit: graph + tuple val(meta), path("SPAdesHybrid-${meta.id}_scaffolds.fasta"), emit: assembly + path "SPAdesHybrid-${meta.id}.log" , emit: log + path "SPAdesHybrid-${meta.id}_contigs.fasta.gz" , emit: contigs_gz + path "SPAdesHybrid-${meta.id}_scaffolds.fasta.gz" , emit: assembly_gz + path "SPAdesHybrid-${meta.id}_graph.gfa.gz" , emit: graph path "versions.yml" , emit: versions script: @@ -30,13 +30,13 @@ process SPADESHYBRID { --pe1-2 ${short_reads[1]} \ --nanopore ${long_reads} \ -o spades - mv spades/assembly_graph_with_scaffolds.gfa ${meta.id}_graph.gfa - mv spades/scaffolds.fasta ${meta.id}_scaffolds.fasta - mv spades/contigs.fasta ${meta.id}_contigs.fasta - mv spades/spades.log ${meta.id}.log - gzip "${meta.id}_contigs.fasta" - gzip "${meta.id}_graph.gfa" - gzip -c "${meta.id}_scaffolds.fasta" > "${meta.id}_scaffolds.fasta.gz" + mv spades/assembly_graph_with_scaffolds.gfa SPAdesHybrid-${meta.id}_graph.gfa + mv spades/scaffolds.fasta SPAdesHybrid-${meta.id}_scaffolds.fasta + mv spades/contigs.fasta SPAdesHybrid-${meta.id}_contigs.fasta + mv spades/spades.log SPAdesHybrid-${meta.id}.log + gzip "SPAdesHybrid-${meta.id}_contigs.fasta" + gzip "SPAdesHybrid-${meta.id}_graph.gfa" + gzip -c "SPAdesHybrid-${meta.id}_scaffolds.fasta" > "SPAdesHybrid-${meta.id}_scaffolds.fasta.gz" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 71fcaafc..edbf26a3 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -33,6 +33,7 @@ workflow BINNING { main: // generate coverage depths for each contig ch_summarizedepth_input = assemblies + .dump(tag: "input_binning") .map { meta, assembly, bams, bais -> def meta_new = meta.clone() [ meta_new, bams, bais ] @@ -47,6 +48,7 @@ workflow BINNING { [ meta_new, depths ] } + .dump(tag: "jgi_depths_out") .set { ch_metabat_depths } // combine depths back with assemblies @@ -61,6 +63,7 @@ workflow BINNING { .map { meta, contigs, reads, indicies, depths -> [ meta, contigs, depths ] } + .dump(tag: "binning_input") // conver metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { @@ -98,9 +101,9 @@ workflow BINNING { // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for // MAG_DEPTHS, first have to separate and re-group due to limitation of // GUNZIP module - METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } - MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } - SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } + METABAT2_METABAT2.out.fasta.transpose().dump(tag: "metabat2_fasta").set { ch_metabat2_results_transposed } + MAXBIN2.out.binned_fastas.transpose().dump(tag: "maxbin2_fasta").set { ch_maxbin2_results_transposed } + SPLIT_FASTA.out.unbinned.transpose().dump(tag: "splitfasta_fasta").set { ch_split_fasta_results_transposed } ch_metabat2_results_transposed .mix( ch_maxbin2_results_transposed, ch_split_fasta_results_transposed ) @@ -138,7 +141,8 @@ workflow BINNING { MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped + METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) + .dump(tag: "final_bins_out") .groupTuple(by: 0) .set{ ch_binning_results_final } diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf index e179e558..bca398dc 100644 --- a/subworkflows/local/binning_preparation.nf +++ b/subworkflows/local/binning_preparation.nf @@ -14,6 +14,7 @@ workflow BINNING_PREPARATION { reads // channel: [ val(meta), [ reads ] ] main: + assemblies.dump(tag: "input_binning_preparation") // build bowtie2 index for all assemblies BOWTIE2_ASSEMBLY_BUILD ( assemblies ) From 98497763d2301798085511151774c079983cf18a Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 8 Mar 2022 14:39:05 +0100 Subject: [PATCH 095/246] More dumps --- subworkflows/local/binning.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index edbf26a3..4b8d8a61 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -84,6 +84,7 @@ workflow BINNING { } if ( !params.skip_maxbin2 ) { + ch_maxbin2_input.dump(tag: "maxbin2_input") MAXBIN2 ( ch_maxbin2_input ) } From ee11868af246fc2a740a4ec27067f27d7dd5f80e Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 15 Mar 2022 15:08:55 +0100 Subject: [PATCH 096/246] Fix binning output files --- modules.json | 2 +- modules/nf-core/modules/maxbin2/main.nf | 3 +- modules/nf-core/modules/maxbin2/meta.yml | 2 +- subworkflows/local/binning.nf | 71 +++++++++++++++-------- subworkflows/local/binning_preparation.nf | 1 - workflows/mag.nf | 19 ++++-- 6 files changed, 63 insertions(+), 35 deletions(-) diff --git a/modules.json b/modules.json index acf7967a..1b1164a9 100644 --- a/modules.json +++ b/modules.json @@ -28,7 +28,7 @@ "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "maxbin2": { - "git_sha": "e20e57f90b6787ac9a010a980cf6ea98bd990046" + "git_sha": "b78a4a456762a4c59fd5023e70f36a27f76d4a97" }, "metabat2/jgisummarizebamcontigdepths": { "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" diff --git a/modules/nf-core/modules/maxbin2/main.nf b/modules/nf-core/modules/maxbin2/main.nf index 7b818d00..a48df43f 100644 --- a/modules/nf-core/modules/maxbin2/main.nf +++ b/modules/nf-core/modules/maxbin2/main.nf @@ -29,8 +29,9 @@ process MAXBIN2 { def prefix = task.ext.prefix ?: "${meta.id}" def associate_files = reads ? "-reads $reads" : "-abund $abund" """ + mkdir input/ && mv $contigs input/ run_MaxBin.pl \\ - -contig $contigs \\ + -contig input/$contigs \\ $associate_files \\ -thread $task.cpus \\ $args \\ diff --git a/modules/nf-core/modules/maxbin2/meta.yml b/modules/nf-core/modules/maxbin2/meta.yml index 358f8323..7971d481 100644 --- a/modules/nf-core/modules/maxbin2/meta.yml +++ b/modules/nf-core/modules/maxbin2/meta.yml @@ -16,7 +16,7 @@ tools: documentation: https://sourceforge.net/projects/maxbin/ tool_dev_url: https://sourceforge.net/projects/maxbin/ doi: "10.1093/bioinformatics/btv638" - licence: ['BSD 3-clause'] + licence: ["BSD 3-clause"] input: - meta: diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 4b8d8a61..af971c28 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -9,7 +9,8 @@ params.mag_depths_summary_options = [:] include { METABAT2_METABAT2 } from '../../modules/nf-core/modules/metabat2/metabat2/main' include { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS } from '../../modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main' include { MAXBIN2 } from '../../modules/nf-core/modules/maxbin2/main' -include { GUNZIP } from '../../modules/nf-core/modules/gunzip/main' +include { GUNZIP as GUNZIP_BINS } from '../../modules/nf-core/modules/gunzip/main' +include { GUNZIP as GUNZIP_UNBINS } from '../../modules/nf-core/modules/gunzip/main' include { CONVERT_DEPTHS } from '../../modules/local/convert_depths' include { SPLIT_FASTA } from '../../modules/local/split_fasta' @@ -31,9 +32,11 @@ workflow BINNING { reads // channel: [ val(meta), [ reads ] ] main: + + ch_versions = Channel.empty() + // generate coverage depths for each contig ch_summarizedepth_input = assemblies - .dump(tag: "input_binning") .map { meta, assembly, bams, bais -> def meta_new = meta.clone() [ meta_new, bams, bais ] @@ -48,9 +51,10 @@ workflow BINNING { [ meta_new, depths ] } - .dump(tag: "jgi_depths_out") .set { ch_metabat_depths } + ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions) + // combine depths back with assemblies ch_metabat2_input = assemblies .map { meta, contigs, reads, indicies -> @@ -63,7 +67,6 @@ workflow BINNING { .map { meta, contigs, reads, indicies, depths -> [ meta, contigs, depths ] } - .dump(tag: "binning_input") // conver metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { @@ -81,11 +84,12 @@ workflow BINNING { // run binning if ( !params.skip_metabat2 ) { METABAT2_METABAT2 ( ch_metabat2_input ) + ch_versions = ch_versions.mix(METABAT2_METABAT2.out.versions) } if ( !params.skip_maxbin2 ) { - ch_maxbin2_input.dump(tag: "maxbin2_input") MAXBIN2 ( ch_maxbin2_input ) + ch_versions = ch_versions.mix(MAXBIN2.out.versions) } // split fastq files, depending @@ -102,32 +106,42 @@ workflow BINNING { // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for // MAG_DEPTHS, first have to separate and re-group due to limitation of // GUNZIP module - METABAT2_METABAT2.out.fasta.transpose().dump(tag: "metabat2_fasta").set { ch_metabat2_results_transposed } - MAXBIN2.out.binned_fastas.transpose().dump(tag: "maxbin2_fasta").set { ch_maxbin2_results_transposed } - SPLIT_FASTA.out.unbinned.transpose().dump(tag: "splitfasta_fasta").set { ch_split_fasta_results_transposed } + METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } + MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } + SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } + ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) + + // mix all bins together ch_metabat2_results_transposed - .mix( ch_maxbin2_results_transposed, ch_split_fasta_results_transposed ) + .mix( ch_maxbin2_results_transposed ) .set { ch_final_bins_for_gunzip } - GUNZIP ( ch_final_bins_for_gunzip ) - GUNZIP.out.gunzip + GUNZIP_BINS ( ch_final_bins_for_gunzip ) + GUNZIP_BINS.out.gunzip .set{ ch_binning_results_gunzipped } + ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions) + + GUNZIP_UNBINS ( ch_split_fasta_results_transposed ) + GUNZIP_UNBINS.out.gunzip + .set{ ch_splitfasta_results_gunzipped } + ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions) // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta for grouping to mix back with original depth // files, as required for MAG_DEPTHS - // Q: where is maxbin2 noclass? All filtered out already? ch_binning_results_gunzipped - .map { meta, results -> - def meta_new = meta.clone() - [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] - } + .mix(ch_splitfasta_results_gunzipped ) + .map { meta, results -> + def meta_new = meta.clone() + [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] + } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) + ch_versions = //ch_versions.mix(MAG_DEPTHS.out.versions) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples @@ -140,20 +154,27 @@ workflow BINNING { MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) + ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.versions).mix(MAG_DEPTHS_SUMMARY.out.versions) // Group final binned contigs per sample for final output + // TODO work out which output channel I want? Have I duplicated? + ch_binning_results_gunzipped + .dump(tag: "final_bins_out_ugz") + .groupTuple(by: 0) + .set{ ch_binning_results_gunzipped_final } + METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) - .dump(tag: "final_bins_out") + .dump(tag: "final_bins_out_gz") .groupTuple(by: 0) - .set{ ch_binning_results_final } + .set{ ch_binning_results_gzipped_final } + + SPLIT_FASTA.out.unbinned.dump(tag: "final_unbins_out_ugz") emit: - bins = ch_binning_results_final - unbinned = SPLIT_FASTA.out.unbinned - tooshort = METABAT2_METABAT2.out.tooshort - lowdepth = METABAT2_METABAT2.out.lowdepth + bins = ch_binning_results_gunzipped_final + bins_gz = ch_binning_results_gzipped_final + unbinned = ch_splitfasta_results_gunzipped.groupTuple() + unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary - metabat2_version = METABAT2_METABAT2.out.versions - metabat2_jgisummarizebamcontigdepths_version = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions - maxbin2_version = MAXBIN2.out.versions + versions = ch_versions } diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf index bca398dc..e179e558 100644 --- a/subworkflows/local/binning_preparation.nf +++ b/subworkflows/local/binning_preparation.nf @@ -14,7 +14,6 @@ workflow BINNING_PREPARATION { reads // channel: [ val(meta), [ reads ] ] main: - assemblies.dump(tag: "input_binning_preparation") // build bowtie2 index for all assemblies BOWTIE2_ASSEMBLY_BUILD ( assemblies ) diff --git a/workflows/mag.nf b/workflows/mag.nf index 2e19e2f1..7a953d6c 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -536,21 +536,19 @@ workflow MAG { ) } - ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) - ch_versions = ch_versions.mix(BINNING.out.metabat2_version.first()) - ch_versions = ch_versions.mix(BINNING.out.metabat2_jgisummarizebamcontigdepths_version.first()) - ch_versions = ch_versions.mix(BINNING.out.maxbin2_version.first()) + //ch_versions = ch_versions.mix(BINNING.out.versions) if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly */ + ch_input_bins_busco = BINNING.out.bins.mix( BINNING.out.unbinned ).transpose().dump(tag: "input_to_busco") BUSCO_QC ( ch_busco_db_file, ch_busco_download_folder, - BINNING.out.bins.transpose() + ch_input_bins_busco ) ch_busco_summary = BUSCO_QC.out.summary ch_busco_multiqc = BUSCO_QC.out.multiqc @@ -564,7 +562,16 @@ workflow MAG { ch_quast_bins_summary = Channel.empty() if (!params.skip_quast){ - QUAST_BINS ( BINNING.out.bins ) + ch_input_for_quast_bins = BINNING.out.bins + .mix( BINNING.out.unbinned ) + .groupTuple() + .map{ + meta, reads -> + def new_reads = reads.flatten() + [meta, new_reads] + } + .dump(tag: "input_for_quast_bins") + QUAST_BINS ( ch_input_for_quast_bins ) ch_versions = ch_versions.mix(QUAST_BINS.out.versions.first()) QUAST_BINS_SUMMARY ( QUAST_BINS.out.quast_bin_summaries.collect() ) ch_quast_bins_summary = QUAST_BINS_SUMMARY.out.summary From 5c6c2205def46754827603e66c59846d91252357 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Tue, 15 Mar 2022 20:58:29 +0000 Subject: [PATCH 097/246] Template update for nf-core/tools version 2.3 --- .github/ISSUE_TEMPLATE/bug_report.yml | 1 - .github/PULL_REQUEST_TEMPLATE.md | 2 +- .github/workflows/awsfulltest.yml | 6 +- .github/workflows/awstest.yml | 6 +- .github/workflows/ci.yml | 4 +- .github/workflows/linting.yml | 28 +-- .gitpod.yml | 14 ++ .nf-core.yml | 1 + .yamllint.yml | 6 + CHANGELOG.md | 2 +- README.md | 6 +- assets/nf-core-mag_logo_light.png | Bin 288 -> 9354 bytes bin/check_samplesheet.py | 346 +++++++++++++++++--------- conf/base.config | 4 +- conf/igenomes.config | 80 +++--- conf/modules.config | 18 +- conf/test.config | 6 +- conf/test_full.config | 6 +- docs/images/nf-core-mag_logo_dark.png | Bin 428 -> 64856 bytes docs/usage.md | 21 +- lib/NfcoreSchema.groovy | 4 +- lib/Utils.groovy | 4 +- lib/WorkflowMag.groovy | 4 +- main.nf | 24 +- nextflow.config | 20 +- nextflow_schema.json | 23 +- subworkflows/local/input_check.nf | 18 +- workflows/mag.nf | 28 +-- 28 files changed, 421 insertions(+), 261 deletions(-) create mode 100644 .gitpod.yml create mode 100644 .nf-core.yml create mode 100644 .yamllint.yml diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml index 8af04d7f..a9dff0f9 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.yml +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -1,4 +1,3 @@ - name: Bug report description: Report something that is broken or incorrect labels: bug diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index d832fa1d..39b8c44a 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -19,7 +19,7 @@ Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/mag/ - [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/mag/tree/master/.github/CONTRIBUTING.md) - [ ] If necessary, also make a PR on the nf-core/mag _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core lint`). -- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). +- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker` --outdir `). - [ ] Usage Documentation in `docs/usage.md` is updated. - [ ] Output Documentation in `docs/output.md` is updated. - [ ] `CHANGELOG.md` is updated. diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 150acfbf..0a02db8f 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -14,7 +14,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@v2 + uses: nf-core/tower-action@v3 # TODO nf-core: You can customise AWS full pipeline tests as required # Add full size test data (but still relatively small datasets for few samples) # on the `test_full.config` test runs with only one set of parameters @@ -31,4 +31,6 @@ jobs: "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-${{ github.sha }}" } profiles: test_full,aws_tower - pre_run_script: 'export NXF_VER=21.10.3' + nextflow_config: | + process.errorStrategy = 'retry' + process.maxRetries = 3 diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index f42d2878..6e009f41 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -11,7 +11,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@v2 + uses: nf-core/tower-action@v3 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} @@ -25,4 +25,6 @@ jobs: "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-test-${{ github.sha }}" } profiles: test,aws_tower - pre_run_script: 'export NXF_VER=21.10.3' + nextflow_config: | + process.errorStrategy = 'retry' + process.maxRetries = 3 diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 59ec52ae..b1c578c1 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -14,7 +14,7 @@ env: jobs: test: - name: Run workflow tests + name: Run pipeline with test data # Only run on push if this is the nf-core dev branch (merged PRs) if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/mag') }} runs-on: ubuntu-latest @@ -47,4 +47,4 @@ jobs: # For example: adding multiple test runs with different parameters # Remember that you can parallelise this by using strategy.matrix run: | - nextflow run ${GITHUB_WORKSPACE} -profile test,docker + nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 3b448773..fda934c0 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -12,9 +12,7 @@ jobs: runs-on: ubuntu-latest steps: - uses: actions/checkout@v2 - - uses: actions/setup-node@v1 - with: - node-version: '10' + - uses: actions/setup-node@v2 - name: Install markdownlint run: npm install -g markdownlint-cli - name: Run Markdownlint @@ -51,9 +49,7 @@ jobs: steps: - uses: actions/checkout@v2 - - uses: actions/setup-node@v1 - with: - node-version: '10' + - uses: actions/setup-node@v2 - name: Install editorconfig-checker run: npm install -g editorconfig-checker @@ -64,14 +60,13 @@ jobs: YAML: runs-on: ubuntu-latest steps: - - uses: actions/checkout@v1 - - uses: actions/setup-node@v1 + - name: Checkout + uses: actions/checkout@master + - name: 'Yamllint' + uses: karancode/yamllint-github-action@master with: - node-version: '10' - - name: Install yaml-lint - run: npm install -g yaml-lint - - name: Run yaml-lint - run: yamllint $(find ${GITHUB_WORKSPACE} -type f -name "*.yml" -o -name "*.yaml") + yamllint_file_or_dir: '.' + yamllint_config_filepath: '.yamllint.yml' # If the above check failed, post a comment on the PR explaining the failure - name: Post PR comment @@ -84,10 +79,11 @@ jobs: To keep the code consistent with lots of contributors, we run automated code consistency checks. To fix this CI test, please run: - * Install `yaml-lint` - * [Install `npm`](https://www.npmjs.com/get-npm) then [install `yaml-lint`](https://www.npmjs.com/package/yaml-lint) (`npm install -g yaml-lint`) + * Install `yamllint` + * Install `yamllint` following [this](https://yamllint.readthedocs.io/en/stable/quickstart.html#installing-yamllint) + instructions or alternative install it in your [conda environment](https://anaconda.org/conda-forge/yamllint) * Fix the markdown errors - * Run the test locally: `yamllint $(find . -type f -name "*.yml" -o -name "*.yaml")` + * Run the test locally: `yamllint $(find . -type f -name "*.yml" -o -name "*.yaml") -c ./.yamllint.yml` * Fix any reported errors in your YAML files Once you push these changes the test should pass, and you can hide this comment :+1: diff --git a/.gitpod.yml b/.gitpod.yml new file mode 100644 index 00000000..b7d4cee1 --- /dev/null +++ b/.gitpod.yml @@ -0,0 +1,14 @@ +image: nfcore/gitpod:latest + +vscode: + extensions: # based on nf-core.nf-core-extensionpack + - codezombiech.gitignore # Language support for .gitignore files + # - cssho.vscode-svgviewer # SVG viewer + - davidanson.vscode-markdownlint # Markdown/CommonMark linting and style checking for Visual Studio Code + - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed + - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files + - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar + - mechatroner.rainbow-csv # Highlight columns in csv files in different colors + # - nextflow.nextflow # Nextflow syntax highlighting + - oderwat.indent-rainbow # Highlight indentation level + - streetsidesoftware.code-spell-checker # Spelling checker for source code diff --git a/.nf-core.yml b/.nf-core.yml new file mode 100644 index 00000000..3805dc81 --- /dev/null +++ b/.nf-core.yml @@ -0,0 +1 @@ +repository_type: pipeline diff --git a/.yamllint.yml b/.yamllint.yml new file mode 100644 index 00000000..d466deec --- /dev/null +++ b/.yamllint.yml @@ -0,0 +1,6 @@ +extends: default + +rules: + document-start: disable + line-length: disable + truthy: disable diff --git a/CHANGELOG.md b/CHANGELOG.md index 2854a2b3..0803027f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v2.1.1 - [date] +## v2.2.0dev - [date] Initial release of nf-core/mag, created with the [nf-core](https://nf-co.re/) template. diff --git a/README.md b/README.md index 7cbbd4d4..39f3d98e 100644 --- a/README.md +++ b/README.md @@ -40,14 +40,14 @@ On release, automated continuous integration tests run the pipeline on a full-si 3. Download the pipeline and test it on a minimal dataset with a single command: ```console - nextflow run nf-core/mag -profile test,YOURPROFILE + nextflow run nf-core/mag -profile test,YOURPROFILE --outdir ``` Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. > * The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. > * Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. - > * If you are using `singularity` and are persistently observing issues downloading Singularity images directly due to timeout or network issues, then you can use the `--singularity_pull_docker_container` parameter to pull and convert the Docker image instead. Alternatively, you can use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. + > * If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. > * If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. 4. Start running your own analysis! @@ -55,7 +55,7 @@ On release, automated continuous integration tests run the pipeline on a full-si ```console - nextflow run nf-core/mag -profile --input samplesheet.csv --genome GRCh37 + nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile ``` ## Documentation diff --git a/assets/nf-core-mag_logo_light.png b/assets/nf-core-mag_logo_light.png index dee3db82069b61e9a5380d940355f47ff565057c..26d7ed5d17ce0345f80ad676dc7da7e928f44ef4 100644 GIT binary patch literal 9354 zcmbt)Wmr^Q)bg&G)oPC#y4Fuu_X{$dp4KLVViORJ$>&71WXLQdLYVTJn`TnkC z_O2;wZ3#4_rEP#?HUrpbG>D8|yu=2|4IQaQMEreMXQrNJek&N%NB)AnzcPWn+9HCM zmPVMdTFJTQy?>2wjVttxxWaf>+~r*JjHjsg?S zEMpy^vj59zQF!iv_)ggmm-^^oUU>LD#{4`z!3xHdfiJ7wQEiZcmPfuGLAyI8V0ED~ z)#m8o55JL}@cZ$6@s-Z8HiSjQb0Ac8j935JTw8stK_?p^5dGA#K+3ub$l?a;i$DbBa_gGc3jH{Bz zYzE&TFbxc!L7)BiP1tm{wwb`Fynd4S32?C;{zsDPuJsc0|8{2M4q^f?!BI7U2wP8- zdpUnW+(2z%i(P{j(B#@IWBo>8RMh862(W=A@L|;Aa;f3K(h8_7>#fQv)tK6rlwdFx z^uJx{ZRf@L@BL`uc<2E#(y0`*=y92%aFB0ob=Hrvo?-phomXe@2tV>aFL;xGtoVr6+c=vg=Zg53;D|ggl4N9 zfFrGc6ljh~-!LvMzGR#RIq_tUCcN@5dY;8I^do#ENlz62GCFJ`VBXini(n|wA-?aZ zM_l{|ZA!D&XlA&tuLij%s1o=_RAYpWoeSd`G`04y%uA<2k^0;)Ta8m|&NMh-3i-rC z@XfwzErf-?UXLVWYdZ#MP-@=Np6M?o4L5a&V z$=`j*G(FdjWOr!2i2Ge>eZ$CgF?Tcef|9W>g-aek^*S!%tSl&VLciE*cX8^?uqzkn zfY@&$47W#hgz|ujxEence51s5E_iy%{}--yyrRT0b@;f7LvQDxE?r4-WP!LV+~MP7 zPH%~x=q^YLyn<{%@>Wr_2wWNqomRL%@8;-yzTy;T?o)c}Flu;N^@iNtOh=#7Ul!|% zc?)R8p=95mh?3jYC2(N?Sz`KKyG{f0!dSZ0WOWKD_KCeQ(BUIC`EiSTmUfmr+h+m` zs|oq_*oNHIj!$yZy-L?@=40;RR_2XoO=p(W1$l=D3lA6bdoJM4jo=L|5bfsmAov&*zJ?rR+$DqkZ!9L=`CWhKf)ls2WF3Y{dO| zE^Ss7s!%7mGZ(~;nw_8vc^Spn_|8zsOcT1P?|SpNZnIjWHO`kbDE?ZiQBKZlRSlx% z={O^`n#kc7vFwKV`ouFV5m_iG;y+!jX(s$GNG|*lwxQ?8`m@9o#K0?~pWLo?59SY& zvPvv1oOAv5nsT1oXCEG1C_A=lRVE7-90`+*%8G+D;cq|sf? 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The order of rows is maintained. + """ -def print_error(error, context="Line", context_str=""): - error_str = "ERROR: Please check samplesheet -> {}".format(error) - if context != "" and context_str != "": - error_str = "ERROR: Please check samplesheet -> {}\n{}: '{}'".format( - error, context.strip(), context_str.strip() + VALID_FORMATS = ( + ".fq.gz", + ".fastq.gz", + ) + + def __init__( + self, + sample_col="sample", + first_col="fastq_1", + second_col="fastq_2", + single_col="single_end", + **kwargs, + ): + """ + Initialize the row checker with the expected column names. + + Args: + sample_col (str): The name of the column that contains the sample name + (default "sample"). + first_col (str): The name of the column that contains the first (or only) + FASTQ file path (default "fastq_1"). + second_col (str): The name of the column that contains the second (if any) + FASTQ file path (default "fastq_2"). + single_col (str): The name of the new column that will be inserted and + records whether the sample contains single- or paired-end sequencing + reads (default "single_end"). + + """ + super().__init__(**kwargs) + self._sample_col = sample_col + self._first_col = first_col + self._second_col = second_col + self._single_col = single_col + self._seen = set() + self.modified = [] + + def validate_and_transform(self, row): + """ + Perform all validations on the given row and insert the read pairing status. + + Args: + row (dict): A mapping from column headers (keys) to elements of that row + (values). + + """ + self._validate_sample(row) + self._validate_first(row) + self._validate_second(row) + self._validate_pair(row) + self._seen.add((row[self._sample_col], row[self._first_col])) + self.modified.append(row) + + def _validate_sample(self, row): + """Assert that the sample name exists and convert spaces to underscores.""" + assert len(row[self._sample_col]) > 0, "Sample input is required." + # Sanitize samples slightly. + row[self._sample_col] = row[self._sample_col].replace(" ", "_") + + def _validate_first(self, row): + """Assert that the first FASTQ entry is non-empty and has the right format.""" + assert len(row[self._first_col]) > 0, "At least the first FASTQ file is required." + self._validate_fastq_format(row[self._first_col]) + + def _validate_second(self, row): + """Assert that the second FASTQ entry has the right format if it exists.""" + if len(row[self._second_col]) > 0: + self._validate_fastq_format(row[self._second_col]) + + def _validate_pair(self, row): + """Assert that read pairs have the same file extension. Report pair status.""" + if row[self._first_col] and row[self._second_col]: + row[self._single_col] = False + assert ( + Path(row[self._first_col]).suffixes == Path(row[self._second_col]).suffixes + ), "FASTQ pairs must have the same file extensions." + else: + row[self._single_col] = True + + def _validate_fastq_format(self, filename): + """Assert that a given filename has one of the expected FASTQ extensions.""" + assert any(filename.endswith(extension) for extension in self.VALID_FORMATS), ( + f"The FASTQ file has an unrecognized extension: {filename}\n" + f"It should be one of: {', '.join(self.VALID_FORMATS)}" ) - print(error_str) - sys.exit(1) + def validate_unique_samples(self): + """ + Assert that the combination of sample name and FASTQ filename is unique. + + In addition to the validation, also rename the sample if more than one sample, + FASTQ file combination exists. + + """ + assert len(self._seen) == len(self.modified), "The pair of sample name and FASTQ must be unique." + if len({pair[0] for pair in self._seen}) < len(self._seen): + counts = Counter(pair[0] for pair in self._seen) + seen = Counter() + for row in self.modified: + sample = row[self._sample_col] + seen[sample] += 1 + if counts[sample] > 1: + row[self._sample_col] = f"{sample}_T{seen[sample]}" + + +def sniff_format(handle): + """ + Detect the tabular format. + + Args: + handle (text file): A handle to a `text file`_ object. The read position is + expected to be at the beginning (index 0). + + Returns: + csv.Dialect: The detected tabular format. + + .. _text file: + https://docs.python.org/3/glossary.html#term-text-file -# TODO nf-core: Update the check_samplesheet function -def check_samplesheet(file_in, file_out): """ - This function checks that the samplesheet follows the following structure: + peek = handle.read(2048) + sniffer = csv.Sniffer() + if not sniffer.has_header(peek): + logger.critical(f"The given sample sheet does not appear to contain a header.") + sys.exit(1) + dialect = sniffer.sniff(peek) + handle.seek(0) + return dialect - sample,fastq_1,fastq_2 - SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz - SAMPLE_PE,SAMPLE_PE_RUN2_1.fastq.gz,SAMPLE_PE_RUN2_2.fastq.gz - SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz, - For an example see: - https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv +def check_samplesheet(file_in, file_out): """ + Check that the tabular samplesheet has the structure expected by nf-core pipelines. - sample_mapping_dict = {} - with open(file_in, "r") as fin: + Validate the general shape of the table, expected columns, and each row. Also add + an additional column which records whether one or two FASTQ reads were found. - ## Check header - MIN_COLS = 2 - # TODO nf-core: Update the column names for the input samplesheet - HEADER = ["sample", "fastq_1", "fastq_2"] - header = [x.strip('"') for x in fin.readline().strip().split(",")] - if header[: len(HEADER)] != HEADER: - print("ERROR: Please check samplesheet header -> {} != {}".format(",".join(header), ",".join(HEADER))) - sys.exit(1) + Args: + file_in (pathlib.Path): The given tabular samplesheet. The format can be either + CSV, TSV, or any other format automatically recognized by ``csv.Sniffer``. + file_out (pathlib.Path): Where the validated and transformed samplesheet should + be created; always in CSV format. - ## Check sample entries - for line in fin: - lspl = [x.strip().strip('"') for x in line.strip().split(",")] - - # Check valid number of columns per row - if len(lspl) < len(HEADER): - print_error( - "Invalid number of columns (minimum = {})!".format(len(HEADER)), - "Line", - line, - ) - num_cols = len([x for x in lspl if x]) - if num_cols < MIN_COLS: - print_error( - "Invalid number of populated columns (minimum = {})!".format(MIN_COLS), - "Line", - line, - ) - - ## Check sample name entries - sample, fastq_1, fastq_2 = lspl[: len(HEADER)] - sample = sample.replace(" ", "_") - if not sample: - print_error("Sample entry has not been specified!", "Line", line) - - ## Check FastQ file extension - for fastq in [fastq_1, fastq_2]: - if fastq: - if fastq.find(" ") != -1: - print_error("FastQ file contains spaces!", "Line", line) - if not fastq.endswith(".fastq.gz") and not fastq.endswith(".fq.gz"): - print_error( - "FastQ file does not have extension '.fastq.gz' or '.fq.gz'!", - "Line", - line, - ) - - ## Auto-detect paired-end/single-end - sample_info = [] ## [single_end, fastq_1, fastq_2] - if sample and fastq_1 and fastq_2: ## Paired-end short reads - sample_info = ["0", fastq_1, fastq_2] - elif sample and fastq_1 and not fastq_2: ## Single-end short reads - sample_info = ["1", fastq_1, fastq_2] - else: - print_error("Invalid combination of columns provided!", "Line", line) - - ## Create sample mapping dictionary = { sample: [ single_end, fastq_1, fastq_2 ] } - if sample not in sample_mapping_dict: - sample_mapping_dict[sample] = [sample_info] - else: - if sample_info in sample_mapping_dict[sample]: - print_error("Samplesheet contains duplicate rows!", "Line", line) - else: - sample_mapping_dict[sample].append(sample_info) - - ## Write validated samplesheet with appropriate columns - if len(sample_mapping_dict) > 0: - out_dir = os.path.dirname(file_out) - make_dir(out_dir) - with open(file_out, "w") as fout: - fout.write(",".join(["sample", "single_end", "fastq_1", "fastq_2"]) + "\n") - for sample in sorted(sample_mapping_dict.keys()): - - ## Check that multiple runs of the same sample are of the same datatype - if not all(x[0] == sample_mapping_dict[sample][0][0] for x in sample_mapping_dict[sample]): - print_error("Multiple runs of a sample must be of the same datatype!", "Sample: {}".format(sample)) - - for idx, val in enumerate(sample_mapping_dict[sample]): - fout.write(",".join(["{}_T{}".format(sample, idx + 1)] + val) + "\n") - else: - print_error("No entries to process!", "Samplesheet: {}".format(file_in)) - - -def main(args=None): - args = parse_args(args) - check_samplesheet(args.FILE_IN, args.FILE_OUT) + Example: + This function checks that the samplesheet follows the following structure, + see also the `viral recon samplesheet`_:: + + sample,fastq_1,fastq_2 + SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz + SAMPLE_PE,SAMPLE_PE_RUN2_1.fastq.gz,SAMPLE_PE_RUN2_2.fastq.gz + SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz, + + .. _viral recon samplesheet: + https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv + + """ + required_columns = {"sample", "fastq_1", "fastq_2"} + # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. + with file_in.open(newline="") as in_handle: + reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle)) + # Validate the existence of the expected header columns. + if not required_columns.issubset(reader.fieldnames): + logger.critical(f"The sample sheet **must** contain the column headers: {', '.join(required_columns)}.") + sys.exit(1) + # Validate each row. + checker = RowChecker() + for i, row in enumerate(reader): + try: + checker.validate_and_transform(row) + except AssertionError as error: + logger.critical(f"{str(error)} On line {i + 2}.") + sys.exit(1) + checker.validate_unique_samples() + header = list(reader.fieldnames) + header.insert(1, "single_end") + # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. + with file_out.open(mode="w", newline="") as out_handle: + writer = csv.DictWriter(out_handle, header, delimiter=",") + writer.writeheader() + for row in checker.modified: + writer.writerow(row) + + +def parse_args(argv=None): + """Define and immediately parse command line arguments.""" + parser = argparse.ArgumentParser( + description="Validate and transform a tabular samplesheet.", + epilog="Example: python check_samplesheet.py samplesheet.csv samplesheet.valid.csv", + ) + parser.add_argument( + "file_in", + metavar="FILE_IN", + type=Path, + help="Tabular input samplesheet in CSV or TSV format.", + ) + parser.add_argument( + "file_out", + metavar="FILE_OUT", + type=Path, + help="Transformed output samplesheet in CSV format.", + ) + parser.add_argument( + "-l", + "--log-level", + help="The desired log level (default WARNING).", + choices=("CRITICAL", "ERROR", "WARNING", "INFO", "DEBUG"), + default="WARNING", + ) + return parser.parse_args(argv) + + +def main(argv=None): + """Coordinate argument parsing and program execution.""" + args = parse_args(argv) + logging.basicConfig(level=args.log_level, format="[%(levelname)s] %(message)s") + if not args.file_in.is_file(): + logger.error(f"The given input file {args.file_in} was not found!") + sys.exit(2) + args.file_out.parent.mkdir(parents=True, exist_ok=True) + check_samplesheet(args.file_in, args.file_out) if __name__ == "__main__": diff --git a/conf/base.config b/conf/base.config index 8ee679ad..a3173752 100644 --- a/conf/base.config +++ b/conf/base.config @@ -1,7 +1,7 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ nf-core/mag Nextflow base config file -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A 'blank slate' config file, appropriate for general use on most high performance compute environments. Assumes that all software is installed and available on the PATH. Runs in `local` mode - all jobs will be run on the logged in environment. diff --git a/conf/igenomes.config b/conf/igenomes.config index 855948de..7a1b3ac6 100644 --- a/conf/igenomes.config +++ b/conf/igenomes.config @@ -1,7 +1,7 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Nextflow config file for iGenomes paths -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Defines reference genomes using iGenome paths. Can be used by any config that customises the base path using: $params.igenomes_base / --igenomes_base @@ -13,7 +13,7 @@ params { genomes { 'GRCh37' { fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/" @@ -26,7 +26,7 @@ params { } 'GRCh38' { fasta = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/" @@ -38,7 +38,7 @@ params { } 'GRCm38' { fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/" @@ -51,7 +51,7 @@ params { } 'TAIR10' { fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/" @@ -62,7 +62,7 @@ params { } 'EB2' { fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/" @@ -72,7 +72,7 @@ params { } 'UMD3.1' { fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/" @@ -83,7 +83,7 @@ params { } 'WBcel235' { fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/" @@ -94,7 +94,7 @@ params { } 'CanFam3.1' { fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/" @@ -105,7 +105,7 @@ params { } 'GRCz10' { fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/" @@ -115,7 +115,7 @@ params { } 'BDGP6' { fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/" @@ -126,7 +126,7 @@ params { } 'EquCab2' { fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/" @@ -137,7 +137,7 @@ params { } 'EB1' { fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/" @@ -147,7 +147,7 @@ params { } 'Galgal4' { fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/" @@ -157,7 +157,7 @@ params { } 'Gm01' { fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/" @@ -167,7 +167,7 @@ params { } 'Mmul_1' { fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/" @@ -178,7 +178,7 @@ params { } 'IRGSP-1.0' { fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/" @@ -188,7 +188,7 @@ params { } 'CHIMP2.1.4' { fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/" @@ -199,7 +199,7 @@ params { } 'Rnor_5.0' { fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BismarkIndex/" @@ -209,7 +209,7 @@ params { } 'Rnor_6.0' { fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/" @@ -219,7 +219,7 @@ params { } 'R64-1-1' { fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/" @@ -230,7 +230,7 @@ params { } 'EF2' { fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/" @@ -242,7 +242,7 @@ params { } 'Sbi1' { fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/" @@ -252,7 +252,7 @@ params { } 'Sscrofa10.2' { fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/" @@ -263,7 +263,7 @@ params { } 'AGPv3' { fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/" @@ -273,7 +273,7 @@ params { } 'hg38' { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/" @@ -285,7 +285,7 @@ params { } 'hg19' { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/" @@ -298,7 +298,7 @@ params { } 'mm10' { fasta = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/" @@ -311,7 +311,7 @@ params { } 'bosTau8' { fasta = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/" @@ -321,7 +321,7 @@ params { } 'ce10' { fasta = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/" @@ -333,7 +333,7 @@ params { } 'canFam3' { fasta = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/" @@ -344,7 +344,7 @@ params { } 'danRer10' { fasta = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/" @@ -355,7 +355,7 @@ params { } 'dm6' { fasta = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/" @@ -366,7 +366,7 @@ params { } 'equCab2' { fasta = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/" @@ -377,7 +377,7 @@ params { } 'galGal4' { fasta = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/" @@ -388,7 +388,7 @@ params { } 'panTro4' { fasta = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/" @@ -399,7 +399,7 @@ params { } 'rn6' { fasta = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/" @@ -409,7 +409,7 @@ params { } 'sacCer3' { fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/" @@ -419,7 +419,7 @@ params { } 'susScr3' { fasta = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa" - bwa = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/version0.6.0/" bowtie2 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/" diff --git a/conf/modules.config b/conf/modules.config index a0506a4d..da58a5d8 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -1,12 +1,12 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Config file for defining DSL2 per module options and publishing paths -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Available keys to override module options: - ext.args = Additional arguments appended to command in module. - ext.args2 = Second set of arguments appended to command in module (multi-tool modules). - ext.args3 = Third set of arguments appended to command in module (multi-tool modules). - ext.prefix = File name prefix for output files. + ext.args = Additional arguments appended to command in module. + ext.args2 = Second set of arguments appended to command in module (multi-tool modules). + ext.args3 = Third set of arguments appended to command in module (multi-tool modules). + ext.prefix = File name prefix for output files. ---------------------------------------------------------------------------------------- */ @@ -14,14 +14,14 @@ process { publishDir = [ path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, - mode: 'copy', + mode: params.publish_dir_mode, saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] withName: SAMPLESHEET_CHECK { publishDir = [ path: { "${params.outdir}/pipeline_info" }, - mode: 'copy', + mode: params.publish_dir_mode, saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] } @@ -33,7 +33,7 @@ process { withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, - mode: 'copy', + mode: params.publish_dir_mode, pattern: '*_versions.yml' ] } diff --git a/conf/test.config b/conf/test.config index 6ce1192d..50d51ccc 100644 --- a/conf/test.config +++ b/conf/test.config @@ -1,11 +1,11 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Nextflow config file for running minimal tests -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test, + nextflow run nf-core/mag -profile test, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_full.config b/conf/test_full.config index 5e0fbbda..a470da07 100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -1,11 +1,11 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Nextflow config file for running full-size tests -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Defines input files and everything required to run a full size pipeline test. 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J-waT~81OO72jb}RoLJM9sUyE8jGGwP=P zT({r52Hx8DE$sF$+szslcT#Se_EpNdtuM&KawTQ+v8aUV!(c1b6Y4w!6D(Cvt4H`E zKADbS@9>1i;=z)V7g#7c)78Q2R21&3KiPRN)L6AW!IagA!AuZn7(F8!n;^O(3&GEL z`IdZS!SoYp3ZUbR4QN~-hc4sVpItBmfoJG&#Lk2yM)p}|n3k}!gi}7^1bED6U@;Ic zl^ka$&sv*cyhCf#Eg<}pQxJ33KjQ@CIv?|QCJpD_6tkR+I~LC|e?kXMA=kc(1G!N4 VJvv>8IrcTl)qtG)%H@^D#V=*Sk$eCE diff --git a/docs/usage.md b/docs/usage.md index db7f656b..0cb8bd3f 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -57,7 +57,7 @@ An [example samplesheet](../assets/samplesheet.csv) has been provided with the p The typical command for running the pipeline is as follows: ```console -nextflow run nf-core/mag --input samplesheet.csv --genome GRCh37 -profile docker +nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile docker ``` This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. @@ -141,11 +141,11 @@ Whilst the default requirements set within the pipeline will hopefully work for For example, if the nf-core/rnaseq pipeline is failing after multiple re-submissions of the `STAR_ALIGN` process due to an exit code of `137` this would indicate that there is an out of memory issue: ```console -[62/149eb0] NOTE: Process `RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) -- Execution is retried (1) -Error executing process > 'RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)' +[62/149eb0] NOTE: Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) -- Execution is retried (1) +Error executing process > 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)' Caused by: - Process `RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) + Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) Command executed: STAR \ @@ -169,17 +169,24 @@ Work dir: Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run` ``` -To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN). We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so based on the search results the file we want is `modules/nf-core/software/star/align/main.nf`. If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9). The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements. The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB. Providing you haven't set any other standard nf-core parameters to __cap__ the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB. The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections. +To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN). +We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so, based on the search results, the file we want is `modules/nf-core/software/star/align/main.nf`. +If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9). +The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements. +The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB. +Providing you haven't set any other standard nf-core parameters to **cap** the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB. +The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections. ```nextflow process { - withName: STAR_ALIGN { + withName: 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN' { memory = 100.GB } } ``` -> **NB:** We specify just the process name i.e. `STAR_ALIGN` in the config file and not the full task name string that is printed to screen in the error message or on the terminal whilst the pipeline is running i.e. `RNASEQ:ALIGN_STAR:STAR_ALIGN`. You may get a warning suggesting that the process selector isn't recognised but you can ignore that if the process name has been specified correctly. This is something that needs to be fixed upstream in core Nextflow. +> **NB:** We specify the full process name i.e. `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN` in the config file because this takes priority over the short name (`STAR_ALIGN`) and allows existing configuration using the full process name to be correctly overridden. +> If you get a warning suggesting that the process selector isn't recognised check that the process name has been specified correctly. ### Updating containers diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy index 40ab65f2..b3d092f8 100755 --- a/lib/NfcoreSchema.groovy +++ b/lib/NfcoreSchema.groovy @@ -27,7 +27,7 @@ class NfcoreSchema { /* groovylint-disable-next-line UnusedPrivateMethodParameter */ public static void validateParameters(workflow, params, log, schema_filename='nextflow_schema.json') { def has_error = false - //=====================================================================// + //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~// // Check for nextflow core params and unexpected params def json = new File(getSchemaPath(workflow, schema_filename=schema_filename)).text def Map schemaParams = (Map) new JsonSlurper().parseText(json).get('definitions') @@ -135,7 +135,7 @@ class NfcoreSchema { } } - //=====================================================================// + //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~// // Validate parameters against the schema InputStream input_stream = new File(getSchemaPath(workflow, schema_filename=schema_filename)).newInputStream() JSONObject raw_schema = new JSONObject(new JSONTokener(input_stream)) diff --git a/lib/Utils.groovy b/lib/Utils.groovy index 1b88aec0..28567bd7 100755 --- a/lib/Utils.groovy +++ b/lib/Utils.groovy @@ -29,12 +29,12 @@ class Utils { conda_check_failed |= !(channels.indexOf('bioconda') < channels.indexOf('defaults')) if (conda_check_failed) { - log.warn "=============================================================================\n" + + log.warn "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + " There is a problem with your Conda configuration!\n\n" + " You will need to set-up the conda-forge and bioconda channels correctly.\n" + " Please refer to https://bioconda.github.io/user/install.html#set-up-channels\n" + " NB: The order of the channels matters!\n" + - "===================================================================================" + "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" } } } diff --git a/lib/WorkflowMag.groovy b/lib/WorkflowMag.groovy index b3a06427..9e3adf54 100755 --- a/lib/WorkflowMag.groovy +++ b/lib/WorkflowMag.groovy @@ -48,11 +48,11 @@ class WorkflowMag { // private static void genomeExistsError(params, log) { if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { - log.error "=============================================================================\n" + + log.error "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + " Genome '${params.genome}' not found in any config files provided to the pipeline.\n" + " Currently, the available genome keys are:\n" + " ${params.genomes.keySet().join(", ")}\n" + - "===================================================================================" + "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" System.exit(1) } } diff --git a/main.nf b/main.nf index 97a6f29d..1bcbb091 100644 --- a/main.nf +++ b/main.nf @@ -1,8 +1,8 @@ #!/usr/bin/env nextflow /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ nf-core/mag -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Github : https://github.com/nf-core/mag Website: https://nf-co.re/mag Slack : https://nfcore.slack.com/channels/mag @@ -12,25 +12,25 @@ nextflow.enable.dsl = 2 /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ GENOME PARAMETER VALUES -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ params.fasta = WorkflowMain.getGenomeAttribute(params, 'fasta') /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ VALIDATE & PRINT PARAMETER SUMMARY -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ WorkflowMain.initialise(workflow, params, log) /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ NAMED WORKFLOW FOR PIPELINE -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ include { MAG } from './workflows/mag' @@ -43,9 +43,9 @@ workflow NFCORE_MAG { } /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ RUN ALL WORKFLOWS -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ // @@ -57,7 +57,7 @@ workflow { } /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ THE END -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ diff --git a/nextflow.config b/nextflow.config index 6a929ee8..50dbaf7e 100644 --- a/nextflow.config +++ b/nextflow.config @@ -1,7 +1,7 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ nf-core/mag Nextflow config file -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Default config options for all compute environments ---------------------------------------------------------------------------------------- */ @@ -24,8 +24,9 @@ params { max_multiqc_email_size = '25.MB' // Boilerplate options - outdir = './results' + outdir = null tracedir = "${params.outdir}/pipeline_info" + publish_dir_mode = 'copy' email = null email_on_fail = null plaintext_email = false @@ -62,6 +63,15 @@ try { System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config") } +// Load nf-core/mag custom profiles from different institutions. +// Warning: Uncomment only if a pipeline-specific instititutional config already exists on nf-core/configs! +// try { +// includeConfig "${params.custom_config_base}/pipeline/mag.config" +// } catch (Exception e) { +// System.err.println("WARNING: Could not load nf-core/config/mag profiles: ${params.custom_config_base}/pipeline/mag.config") +// } + + profiles { debug { process.beforeScript = 'echo $HOSTNAME' } conda { @@ -121,7 +131,7 @@ if (!params.igenomes_ignore) { } // Export these variables to prevent local Python/R libraries from conflicting with those in the container -// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. +// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. // See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable. env { @@ -159,7 +169,7 @@ manifest { description = 'Assembly, binning and annotation of metagenomes' mainScript = 'main.nf' nextflowVersion = '!>=21.10.3' - version = '2.1.1' + version = '2.2.0dev' } // Load modules.config for DSL2 module specific options diff --git a/nextflow_schema.json b/nextflow_schema.json index 22133aa1..bc78304a 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -11,7 +11,8 @@ "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", "required": [ - "input" + "input", + "outdir" ], "properties": { "input": { @@ -26,8 +27,8 @@ }, "outdir": { "type": "string", - "description": "Path to the output directory where the results will be saved.", - "default": "./results", + "format": "directory-path", + "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", "fa_icon": "fas fa-folder-open" }, "email": { @@ -178,6 +179,22 @@ "fa_icon": "fas fa-question-circle", "hidden": true }, + "publish_dir_mode": { + "type": "string", + "default": "copy", + "description": "Method used to save pipeline results to output directory.", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", + "fa_icon": "fas fa-copy", + "enum": [ + "symlink", + "rellink", + "link", + "copy", + "copyNoFollow", + "move" + ], + "hidden": true + }, "email_on_fail": { "type": "string", "description": "Email address for completion summary, only when pipeline fails.", diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index cddcbb3c..0aecf87f 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -12,7 +12,7 @@ workflow INPUT_CHECK { SAMPLESHEET_CHECK ( samplesheet ) .csv .splitCsv ( header:true, sep:',' ) - .map { create_fastq_channels(it) } + .map { create_fastq_channel(it) } .set { reads } emit: @@ -21,22 +21,24 @@ workflow INPUT_CHECK { } // Function to get list of [ meta, [ fastq_1, fastq_2 ] ] -def create_fastq_channels(LinkedHashMap row) { +def create_fastq_channel(LinkedHashMap row) { + // create meta map def meta = [:] - meta.id = row.sample - meta.single_end = row.single_end.toBoolean() + meta.id = row.sample + meta.single_end = row.single_end.toBoolean() - def array = [] + // add path(s) of the fastq file(s) to the meta map + def fastq_meta = [] if (!file(row.fastq_1).exists()) { exit 1, "ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}" } if (meta.single_end) { - array = [ meta, [ file(row.fastq_1) ] ] + fastq_meta = [ meta, [ file(row.fastq_1) ] ] } else { if (!file(row.fastq_2).exists()) { exit 1, "ERROR: Please check input samplesheet -> Read 2 FastQ file does not exist!\n${row.fastq_2}" } - array = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ] + fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ] } - return array + return fastq_meta } diff --git a/workflows/mag.nf b/workflows/mag.nf index a454563b..9dc9e53a 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -1,7 +1,7 @@ /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ VALIDATE INPUTS -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ def summary_params = NfcoreSchema.paramsSummaryMap(workflow, params) @@ -18,18 +18,18 @@ for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input samplesheet not specified!' } /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ CONFIG FILES -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ ch_multiqc_config = file("$projectDir/assets/multiqc_config.yaml", checkIfExists: true) ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multiqc_config) : Channel.empty() /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT LOCAL MODULES/SUBWORKFLOWS -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ // @@ -38,9 +38,9 @@ ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multi include { INPUT_CHECK } from '../subworkflows/local/input_check' /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT NF-CORE MODULES/SUBWORKFLOWS -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ // @@ -51,9 +51,9 @@ include { MULTIQC } from '../modules/nf-core/modules/multiqc include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modules/custom/dumpsoftwareversions/main' /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ RUN MAIN WORKFLOW -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ // Info required for completion email and summary @@ -104,9 +104,9 @@ workflow MAG { } /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ COMPLETION EMAIL AND SUMMARY -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ workflow.onComplete { @@ -117,7 +117,7 @@ workflow.onComplete { } /* -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ THE END -======================================================================================== +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ From 3dfbfed33b66dc28e684aaadef572dcde31e2f39 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 16 Mar 2022 09:42:55 +0100 Subject: [PATCH 098/246] Mix in filter-passted unbins for PROKKA --- workflows/mag.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/mag.nf b/workflows/mag.nf index 7a953d6c..166fd722 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -620,7 +620,7 @@ workflow MAG { /* * Prokka: Genome annotation */ - BINNING.out.bins.transpose() + BINNING.out.bins.mix(BINNING.out.unbinned).transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() From 02e805b4a709f9738253fc1b21c501b3c80fcaca Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 16 Mar 2022 16:47:57 +0100 Subject: [PATCH 099/246] Re activate version reporting --- subworkflows/local/binning.nf | 12 +++++------- workflows/mag.nf | 2 +- 2 files changed, 6 insertions(+), 8 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index af971c28..28871713 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -141,7 +141,7 @@ workflow BINNING { .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) - ch_versions = //ch_versions.mix(MAG_DEPTHS.out.versions) + ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples @@ -154,21 +154,19 @@ workflow BINNING { MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) - ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.versions).mix(MAG_DEPTHS_SUMMARY.out.versions) + ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.versions) + ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions) // Group final binned contigs per sample for final output - // TODO work out which output channel I want? Have I duplicated? ch_binning_results_gunzipped - .dump(tag: "final_bins_out_ugz") .groupTuple(by: 0) .set{ ch_binning_results_gunzipped_final } METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) - .dump(tag: "final_bins_out_gz") .groupTuple(by: 0) .set{ ch_binning_results_gzipped_final } - SPLIT_FASTA.out.unbinned.dump(tag: "final_unbins_out_ugz") + SPLIT_FASTA.out.unbinned emit: bins = ch_binning_results_gunzipped_final @@ -176,5 +174,5 @@ workflow BINNING { unbinned = ch_splitfasta_results_gunzipped.groupTuple() unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary - versions = ch_versions + versions = ch_versions.dump(tag: "binning_versions") } diff --git a/workflows/mag.nf b/workflows/mag.nf index 166fd722..c43d8ee5 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -538,7 +538,7 @@ workflow MAG { ch_bowtie2_assembly_multiqc = BINNING_PREPARATION.out.bowtie2_assembly_multiqc ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) - //ch_versions = ch_versions.mix(BINNING.out.versions) + ch_versions = ch_versions.mix(BINNING.out.versions) if (!params.skip_busco){ /* From 3ab5e07adb26f92e10dbc367c30b48fefbfcd606 Mon Sep 17 00:00:00 2001 From: Paul Cantalupo Date: Wed, 16 Mar 2022 14:33:33 -0400 Subject: [PATCH 100/246] documenting files for Quast --- docs/output.md | 13 +++++++++++-- 1 file changed, 11 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index 12f09260..be29cca0 100644 --- a/docs/output.md +++ b/docs/output.md @@ -209,10 +209,19 @@ SPAdesHybrid is a part of the [SPAdes](http://cab.spbu.ru/software/spades/) soft

Output files -* `Assembly/[assembler]/QC/[sample/group]/` - * `report.*`: QUAST report in various formats, such as html, txt, tsv or tex +* `Assembly/[assembler]/QC/[sample/group]/QUAST/` + * `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt + * `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) * `quast.log`: QUAST log file + * `metaquast.log`: MetaQUAST log file + * `icarus.html`: Icarus main menu with links to interactive viewers + * `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest + * `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) + * `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs + * `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs + * `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. * `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format + * `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor)
From 834f80792c8c971180514414a19b842705397e9e Mon Sep 17 00:00:00 2001 From: Paul Cantalupo Date: Wed, 16 Mar 2022 15:24:24 -0400 Subject: [PATCH 101/246] documenting files for Quast with BINs --- docs/output.md | 13 +++++++++++-- 1 file changed, 11 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index be29cca0..b8ec4841 100644 --- a/docs/output.md +++ b/docs/output.md @@ -299,15 +299,24 @@ For each genome bin the median sequencing depth is computed based on the corresp ### QC for metagenome assembled genomes with QUAST -[QUAST](http://cab.spbu.ru/software/quast/) is a tool that evaluates genome assemblies by computing various metrics. The QUAST output is also included in the MultiQC report, as well as in the assembly directories themselves. +[QUAST](http://cab.spbu.ru/software/quast/) is a tool that evaluates genome assemblies by computing various metrics. The QUAST output is in the bin directories shown below. This QUAST output is not shown in the MultiQC report.
Output files * `GenomeBinning/QC/QUAST/[assembler]-[bin]/` - * `report.*`: QUAST report in various formats, such as html, txt, tsv or tex + * `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt + * `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) * `quast.log`: QUAST log file + * `metaquast.log`: MetaQUAST log file + * `icarus.html`: Icarus main menu with links to interactive viewers + * `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest + * `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) + * `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs + * `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs + * `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. * `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format + * `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor) * `GenomeBinning/QC/` * `quast_summary.tsv`: QUAST output for all bins summarized From d81c2b251839dea7d4d01add7c72981cf88b2790 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 17 Mar 2022 13:07:54 +0100 Subject: [PATCH 102/246] Remove biopython software reporting for MAG_DEPTHS as not used in script, hardcode biopython version for SPLIT_FASTA due to wierd container error --- modules/local/mag_depths.nf | 1 - modules/local/split_fasta.nf | 2 +- 2 files changed, 1 insertion(+), 2 deletions(-) diff --git a/modules/local/mag_depths.nf b/modules/local/mag_depths.nf index e75056be..f013492a 100644 --- a/modules/local/mag_depths.nf +++ b/modules/local/mag_depths.nf @@ -24,7 +24,6 @@ process MAG_DEPTHS { cat <<-END_VERSIONS > versions.yml "${task.process}": python: \$(python --version 2>&1 | sed 's/Python //g') - biopython: \$(biopython --version 2>&1 | sed 's/biopython //g') pandas: \$(python -c "import pkg_resources; print(pkg_resources.get_distribution('pandas').version)") END_VERSIONS """ diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 358e0408..2d27311e 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -27,7 +27,7 @@ process SPLIT_FASTA { cat <<-END_VERSIONS > versions.yml "${task.process}": python: \$(python --version 2>&1 | sed 's/Python //g') - biopython: \$(biopython --version 2>&1 | sed 's/biopython //g') + biopython: 1.7.4 pandas: \$(python -c "import pkg_resources; print(pkg_resources.get_distribution('pandas').version)") END_VERSIONS """ From 5dde80033ef90a3903287d56929e40656d2c4ec1 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 18 Mar 2022 09:38:01 +0100 Subject: [PATCH 103/246] Revert "Merge branch 'dastool' into maxbin" This reverts commit 0ac06b5b52a7979f1cb4fd6b9c2735356105837d, reversing changes made to d81c2b251839dea7d4d01add7c72981cf88b2790. --- conf/modules.config | 11 --- modules.json | 6 -- .../nf-core/modules/dastool/dastool/main.nf | 67 ------------- .../nf-core/modules/dastool/dastool/meta.yml | 99 ------------------- .../modules/dastool/scaffolds2bin/main.nf | 40 -------- .../modules/dastool/scaffolds2bin/meta.yml | 58 ----------- nextflow.config | 7 +- subworkflows/local/binning.nf | 7 -- subworkflows/local/binning_refinement.nf | 76 -------------- workflows/mag.nf | 24 ++--- 10 files changed, 8 insertions(+), 387 deletions(-) delete mode 100644 modules/nf-core/modules/dastool/dastool/main.nf delete mode 100644 modules/nf-core/modules/dastool/dastool/meta.yml delete mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/main.nf delete mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/meta.yml delete mode 100644 subworkflows/local/binning_refinement.nf diff --git a/conf/modules.config b/conf/modules.config index 2a54f24b..7789d405 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -133,7 +133,6 @@ process { filename.indexOf('.contigs.fa.gz') > 0 ? filename : filename.indexOf('.log') > 0 ? filename : null } ] - ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: SPADES { @@ -143,7 +142,6 @@ process { mode: 'copy', pattern: "*.{fasta.gz,gfa.gz,log}" ] - ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: SPADESHYBRID { @@ -153,7 +151,6 @@ process { mode: 'copy', pattern: "*.{fasta.gz,gfa.gz,log}" ] - ext.prefix = { "${meta.assembler}-${meta.id}" } } withName: QUAST { @@ -400,14 +397,6 @@ process { ] } - withName: DASTOOL_SCAFFOLDS2BIN { - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } - } - - withName: DASTOOL_DASTOOL { - ext.prefix = { "${meta.assembler}-${meta.id}" } - } - withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/modules.json b/modules.json index 625570eb..1b1164a9 100644 --- a/modules.json +++ b/modules.json @@ -15,12 +15,6 @@ "custom/dumpsoftwareversions": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, - "dastool/dastool": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" - }, - "dastool/scaffolds2bin": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" - }, "fastp": { "git_sha": "e751e5040af57e1b4e06ed4e0f3efe6de25c1683" }, diff --git a/modules/nf-core/modules/dastool/dastool/main.nf b/modules/nf-core/modules/dastool/dastool/main.nf deleted file mode 100644 index 53dfea19..00000000 --- a/modules/nf-core/modules/dastool/dastool/main.nf +++ /dev/null @@ -1,67 +0,0 @@ -process DASTOOL_DASTOOL { - tag "$meta.id" - label 'process_medium' - - conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : - 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" - - input: - tuple val(meta), path(contigs), path(bins) - path(proteins) - path(db_directory) - val(search_engine) - - output: - tuple val(meta), path("*.log") , emit: log - tuple val(meta), path("*_summary.txt") , emit: summary - tuple val(meta), path("*_DASTool_scaffolds2bin.txt") , emit: scaffolds2bin - tuple val(meta), path("*.eval") , optional: true, emit: eval - tuple val(meta), path("*_DASTool_bins/*.fa") , optional: true, emit: bins - tuple val(meta), path("*.pdf") , optional: true, emit: pdfs - tuple val(meta), path("*.proteins.faa") , optional: true, emit: fasta_proteins - tuple val(meta), path("*.archaea.scg") , optional: true, emit: fasta_archaea_scg - tuple val(meta), path("*.bacteria.scg") , optional: true, emit: fasta_bacteria_scg - tuple val(meta), path("*.seqlength") , optional: true, emit: seqlength - path "versions.yml" , emit: versions - - when: - task.ext.when == null || task.ext.when - - script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - def bin_list = bins instanceof List ? bins.join(",") : "$bins" - def engine = search_engine ? "--search_engine $search_engine" : "--search_engine diamond" - def db_dir = db_directory ? "--db_directory $db_directory" : "" - def clean_contigs = contigs.toString() - ".gz" - def decompress_contigs = contigs.toString() == clean_contigs ? "" : "gunzip -q -f $contigs" - def decompress_proteins = proteins ? "gunzip -f $proteins" : "" - def clean_proteins = proteins ? proteins.toString() - ".gz" : "" - def proteins_pred = proteins ? "--proteins $clean_proteins" : "" - - if (! search_engine) { - log.info('[DAS_Tool] Default search engine (USEARCH) is proprietary software and not available in bioconda. Using DIAMOND as alternative.') - } - - """ - $decompress_proteins - $decompress_contigs - - DAS_Tool \\ - $args \\ - $proteins_pred \\ - $db_dir \\ - $engine \\ - -t $task.cpus \\ - --bins $bin_list \\ - -c $clean_contigs \\ - -o $prefix - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) - END_VERSIONS - """ -} diff --git a/modules/nf-core/modules/dastool/dastool/meta.yml b/modules/nf-core/modules/dastool/dastool/meta.yml deleted file mode 100644 index a77df9bd..00000000 --- a/modules/nf-core/modules/dastool/dastool/meta.yml +++ /dev/null @@ -1,99 +0,0 @@ -name: dastool_dastool -description: DAS Tool binning step. -keywords: - - binning - - das tool - - table - - de novo - - bins - - contigs - - assembly - - das_tool -tools: - - dastool: - description: | - DAS Tool is an automated method that integrates the results - of a flexible number of binning algorithms to calculate an optimized, non-redundant - set of bins from a single assembly. - - homepage: https://github.com/cmks/DAS_Tool - documentation: https://github.com/cmks/DAS_Tool - tool_dev_url: https://github.com/cmks/DAS_Tool - doi: "10.1038/s41564-018-0171-1" - licence: ["BSD"] - -input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - contigs: - type: file - description: fasta file - pattern: "*.{fa.gz,fas.gz,fasta.gz}" - - bins: - type: file - description: "Scaffolds2bin tabular file generated with dastool/scaffolds2bin" - pattern: "*.scaffolds2bin.tsv" - - proteins: - type: file - description: Predicted proteins in prodigal fasta format (>scaffoldID_geneNo) - pattern: "*.{fa.gz,fas.gz,fasta.gz}" - - db_directory: - type: file - description: (optional) Directory of single copy gene database. - - search_engine: - type: val - description: Engine used for single copy gene identification. USEARCH is not supported due to it being proprietary [blast/diamond] - -output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - version: - type: file - description: File containing software version - pattern: "versions.yml" - - log: - type: file - description: Log file of the run - pattern: "*.log" - - summary: - type: file - description: Summary of output bins including quality and completeness estimates - pattern: "*summary.txt" - - scaffolds2bin: - type: file - description: Scaffolds to bin file of output bins - pattern: "*.scaffolds2bin.txt" - - eval: - type: file - description: Quality and completeness estimates of input bin sets - pattern: "*.eval" - - pdfs: - type: file - description: Plots showing the amount of high quality bins and score distribution of bins per method - pattern: "*.pdf" - - fasta_proteins: - type: file - description: Output from prodigal if not already supplied - pattern: "*.proteins.faa" - - fasta_archaea_scg: - type: file - description: Results of archaeal single-copy-gene prediction - pattern: "*.archaea.scg" - - fasta_bacteria_scg: - type: file - description: Results of bacterial single-copy-gene prediction - pattern: "*.bacteria.scg" - - seqlength: - type: file - description: Summary of contig lengths - pattern: "*.seqlength" - -authors: - - "@maxibor" - - "@jfy133" diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf b/modules/nf-core/modules/dastool/scaffolds2bin/main.nf deleted file mode 100644 index 6e9fcdc2..00000000 --- a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf +++ /dev/null @@ -1,40 +0,0 @@ -process DASTOOL_SCAFFOLDS2BIN { - tag "$meta.id" - label 'process_low' - - conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : - 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" - - input: - tuple val(meta), path(fasta) - val(extension) - - output: - tuple val(meta), path("*.tsv"), emit: scaffolds2bin - path "versions.yml" , emit: versions - - when: - task.ext.when == null || task.ext.when - - script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - def file_extension = extension ? extension : "fasta" - - """ - gunzip -f *.${file_extension}.gz - - Fasta_to_Scaffolds2Bin.sh \\ - $args \\ - -i . \\ - -e $file_extension \\ - > ${prefix}.tsv - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) - END_VERSIONS - """ -} diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml b/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml deleted file mode 100644 index 0bf8618d..00000000 --- a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml +++ /dev/null @@ -1,58 +0,0 @@ -name: dastool_scaffolds2bin -description: Helper script to convert a set of bins in fasta format to tabular scaffolds2bin format -keywords: - - binning - - das tool - - table - - de novo - - bins - - contigs - - assembly - - das_tool -tools: - - dastool: - description: | - DAS Tool is an automated method that integrates the results - of a flexible number of binning algorithms to calculate an optimized, non-redundant - set of bins from a single assembly. - - homepage: https://github.com/cmks/DAS_Tool - documentation: https://github.com/cmks/DAS_Tool - tool_dev_url: https://github.com/cmks/DAS_Tool - doi: "10.1038/s41564-018-0171-1" - licence: ["BSD"] - -input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fasta: - type: file - description: Fasta of list of fasta files recommended to be gathered via with .collect() of bins - pattern: "*.{fa,fas,fasta}" - - binner: - type: val - description: Name of the binning software (optional) - - extension: - type: val - description: Fasta file extension (fa | fas | fasta | ...) - -output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" - - scaffolds2bin: - type: file - description: tabular scaffolds2bin file for DAS tool input - pattern: "*.scaffolds2bin.tsv" - -authors: - - "@maxibor" diff --git a/nextflow.config b/nextflow.config index e7dc72ec..d5481c21 100644 --- a/nextflow.config +++ b/nextflow.config @@ -75,11 +75,8 @@ params { lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // binning options - skip_metabat2 = false - skip_maxbin2 = false - - // binning refinement - run_dastool = false + skip_metabat2 = false + skip_maxbin2 = false // Bin QC skip_busco = false diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index ca881a88..28871713 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -157,11 +157,6 @@ workflow BINNING { ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.versions) ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions) - // Final actual bins for bin refinement - METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.fastas) - .groupTuple(by: 0) - .set{ ch_bins_gz } - // Group final binned contigs per sample for final output ch_binning_results_gunzipped .groupTuple(by: 0) @@ -173,8 +168,6 @@ workflow BINNING { SPLIT_FASTA.out.unbinned - - emit: bins = ch_binning_results_gunzipped_final bins_gz = ch_binning_results_gzipped_final diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf deleted file mode 100644 index df070018..00000000 --- a/subworkflows/local/binning_refinement.nf +++ /dev/null @@ -1,76 +0,0 @@ -/* - * Bin refinement with DASTool - */ - -include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' -include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' -include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' - -workflow BINNING_REFINEMENT { - take: - contigs // channel: [ val(meta), path(fasta)) ] - bins // channel: [ val(meta), path(binned_fasta_gz) ] - - main: - ch_versions = Channel.empty() - - // run on each bin file separately due to different suffixes - bins - .dump(tag: "input_to_binrefinement") - .branch{ - maxbin2: it[0]['binner'] == 'MaxBin2' - metabat2: it[0]['binner'] == 'MetaBAT2' - } - .set{ ch_for_scaffolds2bin } - - // generate bin files - if ( !params.skip_metabat2 & !params.skip_maxbin2 ) { - DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_for_scaffolds2bin.metabat2, "fa") - DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_for_scaffolds2bin.maxbin2, "fasta") - DASTOOL_SCAFFOLDS2BIN_METABAT2.out.scaffolds2bin - .mix(DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.scaffolds2bin) - .set { ch_s2b_for_dastool } - ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions, DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions ) - } else if ( !params.skip_metabat2 & params.skip_maxbin2 ){ - DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_for_scaffolds2bin.metabat2, "fa") - DASTOOL_SCAFFOLDS2BIN_METABAT2.out.scaffolds2bin.set { ch_s2b_for_dastool } - ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions ) - } else if ( params.skip_metabat2 & !params.skip_maxbin2 ){ - DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_for_scaffolds2bin.maxbin2, "fasta") - DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.scaffolds2bin.set { ch_s2b_for_dastool } - ch_versions = ch_versions.mix( DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions ) - } - - // combine per sample + assembler combination - // TODO scaffolds2bin remove binner from meta, group - // TODO contigs remove binner from meta, group - // mergin into input for DASTool - ch_s2b_for_dastool - .dump(tag: "s2b_prior_meta_reduc") - .map { meta, results -> - def meta_new = meta.clone() - [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] - } - .dump(tag: "s2b_prior_merge") - .groupTuple() - .set{ ch_s2b_for_join } - - contigs.dump(tag: "input_contigs_to_s2b_join").join(ch_s2b_for_join, by: 0 ).set{ ch_input_for_dastool } - - // TODO now picking up noclass/unbinned? - ch_input_for_dastool.dump(tag: "input_to_dastool") - - // Run DAS_Tool - // TODO offer extra parameters - DASTOOL_DASTOOL( ch_input_for_dastool, [], [], [] ) - - - // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) - - - - emit: - //refined_bins - versions = ch_versions - -} diff --git a/workflows/mag.nf b/workflows/mag.nf index 95c921d5..c43d8ee5 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -86,13 +86,12 @@ include { MULTIQC } from '../modules // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' -include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' -include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' -include { BINNING } from '../subworkflows/local/binning' -include { BINNING_REFINEMENT } from '../subworkflows/local/binning_refinement' -include { BUSCO_QC } from '../subworkflows/local/busco_qc' -include { GTDBTK } from '../subworkflows/local/gtdbtk' +include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' +include { BINNING } from '../subworkflows/local/binning' +include { BUSCO_QC } from '../subworkflows/local/busco_qc' +include { GTDBTK } from '../subworkflows/local/gtdbtk' +include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' /* ======================================================================================== @@ -541,16 +540,6 @@ workflow MAG { ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) ch_versions = ch_versions.mix(BINNING.out.versions) - - /* - * DASTool: Binning refinement - */ - - if ( params.run_dastool ) { - BINNING_REFINEMENT ( ch_assemblies, BINNING.out.bins_gz ) - ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) - } - if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly @@ -647,7 +636,6 @@ workflow MAG { ) ch_versions = ch_versions.mix(PROKKA.out.versions.first()) } - } CUSTOM_DUMPSOFTWAREVERSIONS ( From a268431a00b519ce1f150252153c7a841dc89e86 Mon Sep 17 00:00:00 2001 From: d4straub Date: Tue, 22 Mar 2022 14:19:52 +0100 Subject: [PATCH 104/246] update modules to fix yml linting --- modules.json | 10 +- .../modules/bcftools/consensus/main.nf | 20 ++-- .../modules/bcftools/consensus/meta.yml | 84 ++++++------- modules/nf-core/modules/bcftools/view/main.nf | 11 +- .../nf-core/modules/bcftools/view/meta.yml | 110 +++++++++--------- modules/nf-core/modules/fastqc/main.nf | 3 + modules/nf-core/modules/fastqc/meta.yml | 90 +++++++------- modules/nf-core/modules/gunzip/main.nf | 3 + modules/nf-core/modules/gunzip/meta.yml | 54 ++++----- .../nf-core/modules/samtools/faidx/main.nf | 18 ++- .../nf-core/modules/samtools/faidx/meta.yml | 72 ++++++------ 11 files changed, 248 insertions(+), 227 deletions(-) diff --git a/modules.json b/modules.json index 1b1164a9..4b599336 100644 --- a/modules.json +++ b/modules.json @@ -4,13 +4,13 @@ "repos": { "nf-core/modules": { "bcftools/consensus": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "bcftools/index": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "bcftools/view": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "custom/dumpsoftwareversions": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" @@ -19,13 +19,13 @@ "git_sha": "e751e5040af57e1b4e06ed4e0f3efe6de25c1683" }, "fastqc": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "freebayes": { "git_sha": "7026122ec8696016b0b81496d9d39a7918cdb5b2" }, "gunzip": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "maxbin2": { "git_sha": "b78a4a456762a4c59fd5023e70f36a27f76d4a97" @@ -49,7 +49,7 @@ "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, "samtools/faidx": { - "git_sha": "e0aa89141ffecb5f54d230f7ea46de242b74e084" + "git_sha": "1ad73f1b2abdea9398680d6d20014838135c9a35" } } } diff --git a/modules/nf-core/modules/bcftools/consensus/main.nf b/modules/nf-core/modules/bcftools/consensus/main.nf index 9b9384a6..a0c436e2 100644 --- a/modules/nf-core/modules/bcftools/consensus/main.nf +++ b/modules/nf-core/modules/bcftools/consensus/main.nf @@ -2,10 +2,10 @@ process BCFTOOLS_CONSENSUS { tag "$meta.id" label 'process_medium' - conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) + conda (params.enable_conda ? 'bioconda::bcftools=1.14' : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : - 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" + 'https://depot.galaxyproject.org/singularity/bcftools:1.14--h88f3f91_0' : + 'quay.io/biocontainers/bcftools:1.14--h88f3f91_0' }" input: tuple val(meta), path(vcf), path(tbi), path(fasta) @@ -14,13 +14,19 @@ process BCFTOOLS_CONSENSUS { tuple val(meta), path('*.fa'), emit: fasta path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" """ - cat $fasta | bcftools consensus $vcf $args > ${prefix}.fa - header=\$(head -n 1 ${prefix}.fa | sed 's/>//g') - sed -i 's/\${header}/${meta.id}/g' ${prefix}.fa + cat $fasta \\ + | bcftools \\ + consensus \\ + $vcf \\ + $args \\ + > ${prefix}.fa cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/modules/bcftools/consensus/meta.yml b/modules/nf-core/modules/bcftools/consensus/meta.yml index 761115a6..05a93a56 100644 --- a/modules/nf-core/modules/bcftools/consensus/meta.yml +++ b/modules/nf-core/modules/bcftools/consensus/meta.yml @@ -1,49 +1,49 @@ name: bcftools_consensus description: Compresses VCF files keywords: - - variant calling - - consensus - - VCF + - variant calling + - consensus + - VCF tools: - - consensus: - description: | - Create consensus sequence by applying VCF variants to a reference fasta file. - homepage: http://samtools.github.io/bcftools/bcftools.html - documentation: http://www.htslib.org/doc/bcftools.html - doi: 10.1093/bioinformatics/btp352 - licence: ['MIT'] + - consensus: + description: | + Create consensus sequence by applying VCF variants to a reference fasta file. + homepage: http://samtools.github.io/bcftools/bcftools.html + documentation: http://www.htslib.org/doc/bcftools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - vcf: - type: file - description: VCF file - pattern: "*.{vcf}" - - tbi: - type: file - description: tabix index file - pattern: "*.{tbi}" - - fasta: - type: file - description: FASTA reference file - pattern: "*.{fasta,fa}" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: VCF file + pattern: "*.{vcf}" + - tbi: + type: file + description: tabix index file + pattern: "*.{tbi}" + - fasta: + type: file + description: FASTA reference file + pattern: "*.{fasta,fa}" output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fasta: - type: file - description: FASTA reference consensus file - pattern: "*.{fasta,fa}" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: FASTA reference consensus file + pattern: "*.{fasta,fa}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@joseespinosa" - - "@drpatelh" + - "@joseespinosa" + - "@drpatelh" diff --git a/modules/nf-core/modules/bcftools/view/main.nf b/modules/nf-core/modules/bcftools/view/main.nf index b2cbb580..2a240f4a 100644 --- a/modules/nf-core/modules/bcftools/view/main.nf +++ b/modules/nf-core/modules/bcftools/view/main.nf @@ -2,10 +2,10 @@ process BCFTOOLS_VIEW { tag "$meta.id" label 'process_medium' - conda (params.enable_conda ? "bioconda::bcftools=1.13" : null) + conda (params.enable_conda ? 'bioconda::bcftools=1.14' : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : - 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" + 'https://depot.galaxyproject.org/singularity/bcftools:1.14--h88f3f91_0' : + 'quay.io/biocontainers/bcftools:1.14--h88f3f91_0' }" input: tuple val(meta), path(vcf), path(index) @@ -17,9 +17,12 @@ process BCFTOOLS_VIEW { tuple val(meta), path("*.gz") , emit: vcf path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" def regions_file = regions ? "--regions-file ${regions}" : "" def targets_file = targets ? "--targets-file ${targets}" : "" def samples_file = samples ? "--samples-file ${samples}" : "" diff --git a/modules/nf-core/modules/bcftools/view/meta.yml b/modules/nf-core/modules/bcftools/view/meta.yml index df5b0f8f..326fd1fa 100644 --- a/modules/nf-core/modules/bcftools/view/meta.yml +++ b/modules/nf-core/modules/bcftools/view/meta.yml @@ -1,63 +1,63 @@ name: bcftools_view description: View, subset and filter VCF or BCF files by position and filtering expression. Convert between VCF and BCF keywords: - - variant calling - - view - - bcftools - - VCF + - variant calling + - view + - bcftools + - VCF tools: - - view: - description: | - View, subset and filter VCF or BCF files by position and filtering expression. Convert between VCF and BCF - homepage: http://samtools.github.io/bcftools/bcftools.html - documentation: http://www.htslib.org/doc/bcftools.html - doi: 10.1093/bioinformatics/btp352 - licence: ['MIT'] + - view: + description: | + View, subset and filter VCF or BCF files by position and filtering expression. Convert between VCF and BCF + homepage: http://samtools.github.io/bcftools/bcftools.html + documentation: http://www.htslib.org/doc/bcftools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - vcf: - type: file - description: | - The vcf file to be inspected. - e.g. 'file.vcf' - - index: - type: file - description: | - The tab index for the VCF file to be inspected. - e.g. 'file.tbi' - - regions: - type: file - description: | - Optionally, restrict the operation to regions listed in this file. - e.g. 'file.vcf' - - targets: - type: file - description: | - Optionally, restrict the operation to regions listed in this file (doesn't rely upon index files) - e.g. 'file.vcf' - - samples: - type: file - description: | - Optional, file of sample names to be included or excluded. - e.g. 'file.tsv' + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: | + The vcf file to be inspected. + e.g. 'file.vcf' + - index: + type: file + description: | + The tab index for the VCF file to be inspected. + e.g. 'file.tbi' + - regions: + type: file + description: | + Optionally, restrict the operation to regions listed in this file. + e.g. 'file.vcf' + - targets: + type: file + description: | + Optionally, restrict the operation to regions listed in this file (doesn't rely upon index files) + e.g. 'file.vcf' + - samples: + type: file + description: | + Optional, file of sample names to be included or excluded. + e.g. 'file.tsv' output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - vcf: - type: file - description: VCF normalized output file - pattern: "*.{vcf.gz}" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - vcf: + type: file + description: VCF normalized output file + pattern: "*.{vcf.gz}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@abhi18av" + - "@abhi18av" diff --git a/modules/nf-core/modules/fastqc/main.nf b/modules/nf-core/modules/fastqc/main.nf index d250eca0..ed6b8c50 100644 --- a/modules/nf-core/modules/fastqc/main.nf +++ b/modules/nf-core/modules/fastqc/main.nf @@ -15,6 +15,9 @@ process FASTQC { tuple val(meta), path("*.zip") , emit: zip path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' // Add soft-links to original FastQs for consistent naming in pipeline diff --git a/modules/nf-core/modules/fastqc/meta.yml b/modules/nf-core/modules/fastqc/meta.yml index b09553a3..4da5bb5a 100644 --- a/modules/nf-core/modules/fastqc/meta.yml +++ b/modules/nf-core/modules/fastqc/meta.yml @@ -1,52 +1,52 @@ name: fastqc description: Run FastQC on sequenced reads keywords: - - quality control - - qc - - adapters - - fastq + - quality control + - qc + - adapters + - fastq tools: - - fastqc: - description: | - FastQC gives general quality metrics about your reads. - It provides information about the quality score distribution - across your reads, the per base sequence content (%A/C/G/T). - You get information about adapter contamination and other - overrepresented sequences. - homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ - documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ - licence: ['GPL-2.0-only'] + - fastqc: + description: | + FastQC gives general quality metrics about your reads. + It provides information about the quality score distribution + across your reads, the per base sequence content (%A/C/G/T). + You get information about adapter contamination and other + overrepresented sequences. + homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ + documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ + licence: ["GPL-2.0-only"] input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - reads: - type: file - description: | - List of input FastQ files of size 1 and 2 for single-end and paired-end data, - respectively. + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - html: - type: file - description: FastQC report - pattern: "*_{fastqc.html}" - - zip: - type: file - description: FastQC report archive - pattern: "*_{fastqc.zip}" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - html: + type: file + description: FastQC report + pattern: "*_{fastqc.html}" + - zip: + type: file + description: FastQC report archive + pattern: "*_{fastqc.zip}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@drpatelh" - - "@grst" - - "@ewels" - - "@FelixKrueger" + - "@drpatelh" + - "@grst" + - "@ewels" + - "@FelixKrueger" diff --git a/modules/nf-core/modules/gunzip/main.nf b/modules/nf-core/modules/gunzip/main.nf index 77a4e546..9d4b0666 100644 --- a/modules/nf-core/modules/gunzip/main.nf +++ b/modules/nf-core/modules/gunzip/main.nf @@ -14,6 +14,9 @@ process GUNZIP { tuple val(meta), path("$gunzip"), emit: gunzip path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' gunzip = archive.toString() - '.gz' diff --git a/modules/nf-core/modules/gunzip/meta.yml b/modules/nf-core/modules/gunzip/meta.yml index ea1f1546..4d2ebc84 100644 --- a/modules/nf-core/modules/gunzip/meta.yml +++ b/modules/nf-core/modules/gunzip/meta.yml @@ -1,34 +1,34 @@ name: gunzip description: Compresses and decompresses files. keywords: - - gunzip - - compression + - gunzip + - compression tools: - - gunzip: - description: | - gzip is a file format and a software application used for file compression and decompression. - documentation: https://www.gnu.org/software/gzip/manual/gzip.html - licence: ['GPL-3.0-or-later'] + - gunzip: + description: | + gzip is a file format and a software application used for file compression and decompression. + documentation: https://www.gnu.org/software/gzip/manual/gzip.html + licence: ["GPL-3.0-or-later"] input: - - meta: - type: map - description: | - Optional groovy Map containing meta information - e.g. [ id:'test', single_end:false ] - - archive: - type: file - description: File to be compressed/uncompressed - pattern: "*.*" + - meta: + type: map + description: | + Optional groovy Map containing meta information + e.g. [ id:'test', single_end:false ] + - archive: + type: file + description: File to be compressed/uncompressed + pattern: "*.*" output: - - gunzip: - type: file - description: Compressed/uncompressed file - pattern: "*.*" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - gunzip: + type: file + description: Compressed/uncompressed file + pattern: "*.*" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@joseespinosa" - - "@drpatelh" - - "@jfy133" + - "@joseespinosa" + - "@drpatelh" + - "@jfy133" diff --git a/modules/nf-core/modules/samtools/faidx/main.nf b/modules/nf-core/modules/samtools/faidx/main.nf index d8308b03..7732a4ec 100644 --- a/modules/nf-core/modules/samtools/faidx/main.nf +++ b/modules/nf-core/modules/samtools/faidx/main.nf @@ -2,22 +2,28 @@ process SAMTOOLS_FAIDX { tag "$fasta" label 'process_low' - conda (params.enable_conda ? "bioconda::samtools=1.14" : null) + conda (params.enable_conda ? "bioconda::samtools=1.15" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/samtools:1.14--hb421002_0' : - 'quay.io/biocontainers/samtools:1.14--hb421002_0' }" + 'https://depot.galaxyproject.org/singularity/samtools:1.15--h1170115_1' : + 'quay.io/biocontainers/samtools:1.15--h1170115_1' }" input: tuple val(meta), path(fasta) output: - tuple val(meta), path ("*.fai") , emit: fai - path "versions.yml" , emit: versions + tuple val(meta), path ("*.fai"), emit: fai + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when script: def args = task.ext.args ?: '' """ - samtools faidx $fasta + samtools \\ + faidx \\ + $fasta + cat <<-END_VERSIONS > versions.yml "${task.process}": samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') diff --git a/modules/nf-core/modules/samtools/faidx/meta.yml b/modules/nf-core/modules/samtools/faidx/meta.yml index bae97a39..e9767764 100644 --- a/modules/nf-core/modules/samtools/faidx/meta.yml +++ b/modules/nf-core/modules/samtools/faidx/meta.yml @@ -1,43 +1,43 @@ name: samtools_faidx description: Index FASTA file keywords: - - index - - fasta + - index + - fasta tools: - - samtools: - description: | - SAMtools is a set of utilities for interacting with and post-processing - short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. - These files are generated as output by short read aligners like BWA. - homepage: http://www.htslib.org/ - documentation: http://www.htslib.org/doc/samtools.html - doi: 10.1093/bioinformatics/btp352 - licence: ['MIT'] + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fasta: - type: file - description: FASTA file - pattern: "*.{fa,fasta}" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: FASTA file + pattern: "*.{fa,fasta}" output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fai: - type: file - description: FASTA index file - pattern: "*.{fai}" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fai: + type: file + description: FASTA index file + pattern: "*.{fai}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@drpatelh" - - "@ewels" - - "@phue" + - "@drpatelh" + - "@ewels" + - "@phue" From 820c40f33c4113d03a9052f1f4aac8db9eeceee2 Mon Sep 17 00:00:00 2001 From: d4straub Date: Tue, 22 Mar 2022 14:41:05 +0100 Subject: [PATCH 105/246] fix linting --- modules/local/convert_depths.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/convert_depths.nf b/modules/local/convert_depths.nf index 1f5258bf..dcebe79a 100644 --- a/modules/local/convert_depths.nf +++ b/modules/local/convert_depths.nf @@ -17,7 +17,7 @@ process CONVERT_DEPTHS { def prefix = task.ext.prefix ?: "${meta.id}" """ gunzip $depth - bioawk -t '{{if (NR > 1){{print \$1, \$3}}}}' ${depth.toString() - '.gz'} > ${prefix}_mb2_depth.txt + bioawk -t '{ { if (NR > 1) { { print \$1, \$3 } } } }' ${depth.toString() - '.gz'} > ${prefix}_mb2_depth.txt cat <<-END_VERSIONS > versions.yml "${task.process}": From 5f7887aea792be98a7c9d62c25424c0768a44269 Mon Sep 17 00:00:00 2001 From: d4straub Date: Tue, 22 Mar 2022 14:46:28 +0100 Subject: [PATCH 106/246] update CHANGELOG --- CHANGELOG.md | 7 +++++++ 1 file changed, 7 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index ee468daa..e2579dea 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,6 +17,13 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Fixed` +### `Dependencies` + +| Tool | Previous version | New version | +| --- | --- | --- | +| bcftools | 1.13 | 1.14 | +| samtools | 1.14 | 1.15 | + ## v2.1.1 - 2021/11/25 ### `Added` From 6148bdedf8d99ae3a0248d3d5194e15aae3957f5 Mon Sep 17 00:00:00 2001 From: d4straub Date: Tue, 22 Mar 2022 16:55:11 +0100 Subject: [PATCH 107/246] fix BCFTOOLS_VIEW --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index aaba4d91..298a37a9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -282,7 +282,7 @@ process { withName: BCFTOOLS_VIEW { ext.args = "-v snps,mnps -i 'QUAL>=${params.bcftools_view_high_variant_quality} || (QUAL>=${params.bcftools_view_medium_variant_quality} && FORMAT/AO>=${params.bcftools_view_minimal_allelesupport})'" - ext.suffix = ".filtered" + ext.prefix = { "${meta.id}.filtered" } publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, pattern: "*.vcf.gz" From 21636c6d5bbff5093c73e9094133310da3d2d265 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 23 Mar 2022 08:45:48 +0100 Subject: [PATCH 108/246] Add AdapterRemoval2 as an alternative trimmer --- CHANGELOG.md | 1 + modules.json | 3 + modules/local/multiqc.nf | 2 +- .../nf-core/modules/adapterremoval/main.nf | 70 +++++++++++++++ .../nf-core/modules/adapterremoval/meta.yml | 90 +++++++++++++++++++ nextflow.config | 3 +- subworkflows/local/binning.nf | 2 +- workflows/mag.nf | 75 ++++++++++++---- 8 files changed, 226 insertions(+), 20 deletions(-) create mode 100644 modules/nf-core/modules/adapterremoval/main.nf create mode 100644 modules/nf-core/modules/adapterremoval/meta.yml diff --git a/CHANGELOG.md b/CHANGELOG.md index 96f94b7e..2b583685 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,6 +10,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool +- [#264](https://github.com/nf-core/mag/issues/284) - Add AdapterRemoval2 as an alternative read trimmer ### `Changed` diff --git a/modules.json b/modules.json index 1b1164a9..b9a2bb74 100644 --- a/modules.json +++ b/modules.json @@ -3,6 +3,9 @@ "homePage": "https://github.com/nf-core/mag", "repos": { "nf-core/modules": { + "adapterremoval": { + "git_sha": "f0800157544a82ae222931764483331a81812012" + }, "bcftools/consensus": { "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" }, diff --git a/modules/local/multiqc.nf b/modules/local/multiqc.nf index 5faa7a4f..778eb97f 100644 --- a/modules/local/multiqc.nf +++ b/modules/local/multiqc.nf @@ -10,12 +10,12 @@ process MULTIQC { path multiqc_files path mqc_custom_config path 'fastqc_raw/*' - path 'fastp/*' path 'fastqc_trimmed/*' path host_removal path 'quast*/*' path 'bowtie2log/*' path short_summary + path additional output: path "*multiqc_report.html", emit: report diff --git a/modules/nf-core/modules/adapterremoval/main.nf b/modules/nf-core/modules/adapterremoval/main.nf new file mode 100644 index 00000000..9d16b9c9 --- /dev/null +++ b/modules/nf-core/modules/adapterremoval/main.nf @@ -0,0 +1,70 @@ +process ADAPTERREMOVAL { + tag "$meta.id" + label 'process_medium' + + conda (params.enable_conda ? "bioconda::adapterremoval=2.3.2" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/adapterremoval:2.3.2--hb7ba0dd_0' : + 'quay.io/biocontainers/adapterremoval:2.3.2--hb7ba0dd_0' }" + + input: + tuple val(meta), path(reads) + path(adapterlist) + + output: + tuple val(meta), path("${prefix}.truncated.gz") , optional: true, emit: singles_truncated + tuple val(meta), path("${prefix}.discarded.gz") , optional: true, emit: discarded + tuple val(meta), path("${prefix}.pair1.truncated.gz") , optional: true, emit: pair1_truncated + tuple val(meta), path("${prefix}.pair2.truncated.gz") , optional: true, emit: pair2_truncated + tuple val(meta), path("${prefix}.collapsed.gz") , optional: true, emit: collapsed + tuple val(meta), path("${prefix}.collapsed.truncated.gz") , optional: true, emit: collapsed_truncated + tuple val(meta), path("${prefix}.paired.gz") , optional: true, emit: paired_interleaved + tuple val(meta), path('*.log') , emit: log + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def list = adapterlist ? "--adapter-list ${adapterlist}" : "" + prefix = task.ext.prefix ?: "${meta.id}" + + if (meta.single_end) { + """ + AdapterRemoval \\ + --file1 $reads \\ + $args \\ + $adapterlist \\ + --basename ${prefix} \\ + --threads ${task.cpus} \\ + --settings ${prefix}.log \\ + --seed 42 \\ + --gzip + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + adapterremoval: \$(AdapterRemoval --version 2>&1 | sed -e "s/AdapterRemoval ver. //g") + END_VERSIONS + """ + } else { + """ + AdapterRemoval \\ + --file1 ${reads[0]} \\ + --file2 ${reads[1]} \\ + $args \\ + $adapterlist \\ + --basename ${prefix} \\ + --threads $task.cpus \\ + --settings ${prefix}.log \\ + --seed 42 \\ + --gzip + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + adapterremoval: \$(AdapterRemoval --version 2>&1 | sed -e "s/AdapterRemoval ver. //g") + END_VERSIONS + """ + } + +} diff --git a/modules/nf-core/modules/adapterremoval/meta.yml b/modules/nf-core/modules/adapterremoval/meta.yml new file mode 100644 index 00000000..5faad043 --- /dev/null +++ b/modules/nf-core/modules/adapterremoval/meta.yml @@ -0,0 +1,90 @@ +name: adapterremoval +description: Trim sequencing adapters and collapse overlapping reads +keywords: + - trimming + - adapters + - merging + - fastq +tools: + - adapterremoval: + description: The AdapterRemoval v2 tool for merging and clipping reads. + homepage: https://github.com/MikkelSchubert/adapterremoval + documentation: https://adapterremoval.readthedocs.io + licence: ["GPL v3"] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. + pattern: "*.{fq,fastq,fq.gz,fastq.gz}" + - adapterlist: + type: file + description: Optional text file containing list of adapters to look for for removal + with one adapter per line. Otherwise will look for default adapters (see + AdapterRemoval man page), or can be modified to remove user-specified + adapters via ext.args. + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - singles_truncated: + type: file + description: | + Adapter trimmed FastQ files of either single-end reads, or singleton + 'orphaned' reads from merging of paired-end data (i.e., one of the pair + was lost due to filtering thresholds). + pattern: "*.truncated.gz" + - discarded: + type: file + description: | + Adapter trimmed FastQ files of reads that did not pass filtering + thresholds. + pattern: "*.discarded.gz" + - pair1_truncated: + type: file + description: | + Adapter trimmed R1 FastQ files of paired-end reads that did not merge + with their respective R2 pair due to long templates. The respective pair + is stored in 'pair2_truncated'. + pattern: "*.pair1.truncated.gz" + - pair2_truncated: + type: file + description: | + Adapter trimmed R2 FastQ files of paired-end reads that did not merge + with their respective R1 pair due to long templates. The respective pair + is stored in 'pair1_truncated'. + pattern: "*.pair2.truncated.gz" + - collapsed: + type: file + description: | + Collapsed FastQ of paired-end reads that successfully merged with their + respective R1 pair but were not trimmed. + pattern: "*.collapsed.gz" + - collapsed_truncated: + type: file + description: | + Collapsed FastQ of paired-end reads that successfully merged with their + respective R1 pair and were trimmed of adapter due to sufficient overlap. + pattern: "*.collapsed.truncated.gz" + - log: + type: file + description: AdapterRemoval log file + pattern: "*.log" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + +authors: + - "@maxibor" + - "@jfy133" diff --git a/nextflow.config b/nextflow.config index 88f83e6c..e3b67f64 100644 --- a/nextflow.config +++ b/nextflow.config @@ -14,11 +14,12 @@ params { single_end = false // short read preprocessing options + clip_tool = 'fastp' save_trimmed_fail = false fastp_qualified_quality = 15 fastp_cut_mean_quality = 15 keep_phix = false - // phix_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Enterobacteria_phage_phiX174_sensu_lato/all_assembly_versions/GCA_002596845.1_ASM259684v1/GCA_002596845.1_ASM259684v1_genomic.fna.gz" + // phix_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Enterobacteria_phage_phiX174_sensu_lato/all_assembly_versions/GCA_002596845.1_ASM259684v1/GCA_002596845.1_ASM259684v1_genomic.fna.gz" phix_reference = "${baseDir}/assets/data/GCA_002596845.1_ASM259684v1_genomic.fna.gz" host_fasta = null host_genome = null diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 28871713..6ba4b02d 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -174,5 +174,5 @@ workflow BINNING { unbinned = ch_splitfasta_results_gunzipped.groupTuple() unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary - versions = ch_versions.dump(tag: "binning_versions") + versions = ch_versions } diff --git a/workflows/mag.nf b/workflows/mag.nf index c43d8ee5..32dbbb17 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -102,12 +102,14 @@ include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_d // // MODULE: Installed directly from nf-core/modules // -include { FASTQC as FASTQC_RAW } from '../modules/nf-core/modules/fastqc/main' -include { FASTQC as FASTQC_TRIMMED } from '../modules/nf-core/modules/fastqc/main' -include { FASTP } from '../modules/nf-core/modules/fastp/main' -include { PRODIGAL } from '../modules/nf-core/modules/prodigal/main' -include { PROKKA } from '../modules/nf-core/modules/prokka/main' -include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modules/custom/dumpsoftwareversions/main' +include { FASTQC as FASTQC_RAW } from '../modules/nf-core/modules/fastqc/main' +include { FASTQC as FASTQC_TRIMMED } from '../modules/nf-core/modules/fastqc/main' +include { FASTP } from '../modules/nf-core/modules/fastp/main' +include { ADAPTERREMOVAL as ADAPTERREMOVAL_PE } from '../modules/nf-core/modules/adapterremoval/main' +include { ADAPTERREMOVAL as ADAPTERREMOVAL_SE } from '../modules/nf-core/modules/adapterremoval/main' +include { PRODIGAL } from '../modules/nf-core/modules/prodigal/main' +include { PROKKA } from '../modules/nf-core/modules/prokka/main' +include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modules/custom/dumpsoftwareversions/main' //////////////////////////////////////////////////// /* -- Create channel for reference databases -- */ @@ -223,13 +225,45 @@ workflow MAG { ) ch_versions = ch_versions.mix(FASTQC_RAW.out.versions.first()) - FASTP ( - ch_raw_short_reads, - [], - [] - ) - ch_short_reads = FASTP.out.reads - ch_versions = ch_versions.mix(FASTP.out.versions.first()) + if ( params.clip_tool == 'fastp' ) { + ch_clipmerge_out = FASTP ( + ch_raw_short_reads, + [], + [] + ) + ch_short_reads = FASTP.out.reads + ch_versions = ch_versions.mix(FASTP.out.versions.first()) + + } else if ( params.clip_tool == 'adapterremoval' ) { + + // due to strange output file scheme in AR2, have to manually separate + // SE/PE to allow correct pulling of reads after. + ch_adapterremoval_in = ch_raw_short_reads + .branch { + single: it[0]['single_end'] + paired: !it[0]['single_end'] + } + + ADAPTERREMOVAL_PE ( ch_adapterremoval_in.paired, [] ) + ADAPTERREMOVAL_SE ( ch_adapterremoval_in.single, [] ) + + // pair1 and 2 come for PE data from separate output channels, so bring + // this back together again here + ch_adapterremoval_pe_out = Channel.empty() + ch_adapterremoval_pe_out = ADAPTERREMOVAL_PE.out.pair1_truncated + .join(ADAPTERREMOVAL_PE.out.pair2_truncated) + .dump(tag: "ar_pe_mix_out") + .map { + [ it[0], [it[1], it[2]] ] + } + .dump(tag: "ar2_pe_out") + + ch_short_reads = Channel.empty() + ch_short_reads = ch_short_reads.mix(ADAPTERREMOVAL_SE.out.singles_truncated, ch_adapterremoval_pe_out) + + ch_versions = ch_versions.mix(ADAPTERREMOVAL_PE.out.versions, ADAPTERREMOVAL_SE.out.versions) + + } if (params.host_fasta){ BOWTIE2_HOST_REMOVAL_BUILD ( @@ -544,7 +578,7 @@ workflow MAG { /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly */ - ch_input_bins_busco = BINNING.out.bins.mix( BINNING.out.unbinned ).transpose().dump(tag: "input_to_busco") + ch_input_bins_busco = BINNING.out.bins.mix( BINNING.out.unbinned ).transpose() BUSCO_QC ( ch_busco_db_file, ch_busco_download_folder, @@ -570,7 +604,6 @@ workflow MAG { def new_reads = reads.flatten() [meta, new_reads] } - .dump(tag: "input_for_quast_bins") QUAST_BINS ( ch_input_for_quast_bins ) ch_versions = ch_versions.mix(QUAST_BINS.out.versions.first()) QUAST_BINS_SUMMARY ( QUAST_BINS.out.quast_bin_summaries.collect() ) @@ -667,16 +700,24 @@ workflow MAG { */ //This is the local module because (1) fastq file name clashes [probably solveable] and (2) host removal bowtie reporting + ch_multiqc_additional = Channel.empty() + + if ( params.clip_tool == "fastp") { + ch_multiqc_additional = ch_multiqc_additional.mix(FASTP.out.json.collect{it[1]}.ifEmpty([])) + } else if ( params.clip_tool == "adapterremoval" ) { + ch_multiqc_additional = ch_multiqc_additional.mix(ADAPTERREMOVAL_PE.out.log.collect{it[1]}.ifEmpty([]), ADAPTERREMOVAL_SE.out.log.collect{it[1]}.ifEmpty([])) + } + MULTIQC ( ch_multiqc_files.collect(), ch_multiqc_custom_config.collect().ifEmpty([]), FASTQC_RAW.out.zip.collect{it[1]}.ifEmpty([]), - FASTP.out.json.collect{it[1]}.ifEmpty([]), FASTQC_TRIMMED.out.zip.collect{it[1]}.ifEmpty([]), ch_bowtie2_removal_host_multiqc.collect{it[1]}.ifEmpty([]), ch_quast_multiqc.collect().ifEmpty([]), ch_bowtie2_assembly_multiqc.collect().ifEmpty([]), - ch_busco_multiqc.collect().ifEmpty([]) + ch_busco_multiqc.collect().ifEmpty([]), + ch_multiqc_additional ) multiqc_report = MULTIQC.out.report.toList() ch_versions = ch_versions.mix(MULTIQC.out.versions) From 1959af760e057e89b77f7744c984e89e53bdd433 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 23 Mar 2022 14:36:04 +0100 Subject: [PATCH 109/246] Add AdapterRemoval2 options and docs --- CHANGELOG.md | 2 +- assets/multiqc_config.yaml | 5 +++++ conf/modules.config | 33 ++++++++++++++++++++++++++- docs/output.md | 14 ++++++++++++ modules/local/multiqc.nf | 6 ++--- nextflow.config | 7 +++++- nextflow_schema.json | 46 ++++++++++++++++++++++++++++++++------ workflows/mag.nf | 4 ++-- 8 files changed, 102 insertions(+), 15 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 2b583685..d916cd96 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,7 +10,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool -- [#264](https://github.com/nf-core/mag/issues/284) - Add AdapterRemoval2 as an alternative read trimmer +- [#264](https://github.com/nf-core/mag/pull/265) - Add AdapterRemoval2 as an alternative read trimmer ### `Changed` diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml index 8c0214e3..65639f09 100644 --- a/assets/multiqc_config.yaml +++ b/assets/multiqc_config.yaml @@ -17,6 +17,8 @@ top_modules: name: 'FastQC: raw reads' path_filters_exclude: - '*trimmed*' +- 'fastp' +- 'adapterRemoval': - custom_content - 'fastqc': name: 'FastQC: after preprocessing' @@ -42,6 +44,9 @@ custom_data: sp: host_removal: fn: 'host_removal_metrics.tsv' + adapterRemoval: + fn: '*_ar2_*.log' extra_fn_clean_exts: - '.bowtie2' + - '_ar2' diff --git a/conf/modules.config b/conf/modules.config index 7b3bfc49..5566ed6b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -34,7 +34,8 @@ process { "-q ${params.fastp_qualified_quality}", "--cut_front", "--cut_tail", - "--cut_mean_quality ${params.fastp_cut_mean_quality}" + "--cut_mean_quality ${params.fastp_cut_mean_quality}", + "--length_required ${params.reads_minlength} " ].join(' ').trim() publishDir = [ path: { "${params.outdir}/QC_shortreads/fastp/${meta.id}" }, @@ -43,6 +44,36 @@ process { ] } + withName: ADAPTERREMOVAL_PE { + ext.args = [ + "--minlength ${params.reads_minlength}", + "--adapter1 ${params.adapterremoval_adapter1} --adapter2 ${params.adapterremoval_adapter2}", + "--minquality ${params.adapterremoval_minquality} --trimns", + params.adapterremoval_trim_quality_stretch ? "--trim_qualities" : "--trimwindows 4" + ].join(' ').trim() + publishDir = [ + path: { "${params.outdir}/QC_shortreads/adapterremoval/${meta.id}" }, + mode: 'copy', + pattern: "*.{log}" + ] + ext.prefix = { "${meta.id}_ar2" } + } + + withName: ADAPTERREMOVAL_SE { + ext.args = [ + "--minlength ${params.reads_minlength}", + "--adapter1 ${params.adapterremoval_adapter1}", + "--minquality ${params.adapterremoval_minquality} --trimns", + params.adapterremoval_trim_quality_stretch ? "--trim_qualities" : "--trimwindows 4" + ].join(' ').trim() + publishDir = [ + path: { "${params.outdir}/QC_shortreads/adapterremoval/${meta.id}" }, + mode: 'copy', + pattern: "*.{log}" + ] + ext.prefix = { "${meta.id}_ar2" } + } + withName: BOWTIE2_PHIX_REMOVAL_ALIGN { ext.suffix = "phix_removed" publishDir = [ diff --git a/docs/output.md b/docs/output.md index a389e697..e29274e4 100644 --- a/docs/output.md +++ b/docs/output.md @@ -57,6 +57,20 @@ FastQC is run for visualising the general quality metrics of the sequencing runs
+### AdapterRemoval2 + +[fastp](https://github.com/OpenGene/fastp) is a all-in-one fastq preprocessor for read/adapter trimming and quality control. It is used in this pipeline for trimming adapter sequences and discard low-quality reads. Its output is in the results folder and part of the MultiQC report. + +[AdapterRemoval](https://adapterremoval.readthedocs.io/en/stable/) searches for and removes remnant adapter sequences from High-Throughput Sequencing (HTS) data and (optionally) trims low quality bases from the 3' end of reads following adapter removal. It is popular in the field of palaeogenomics. The output logs are stored in the results folder, and as a part of the MultiQC report. + +
+Output files + +* `QC_shortreads/adapterremoval/[sample]/` + * `[sample]_ar2.log`: AdapterRemoval log file (normally called `.settings` by AdapterRemoval.) + +
+ ### Remove PhiX sequences from short reads The pipeline uses bowtie2 to map the reads against PhiX and removes mapped reads. diff --git a/modules/local/multiqc.nf b/modules/local/multiqc.nf index 778eb97f..6d8b2cbb 100644 --- a/modules/local/multiqc.nf +++ b/modules/local/multiqc.nf @@ -1,10 +1,10 @@ process MULTIQC { label 'process_medium' - conda (params.enable_conda ? "bioconda::multiqc=1.11" : null) + conda (params.enable_conda ? "bioconda::multiqc=1.12" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/multiqc:1.11--pyhdfd78af_0' : - 'quay.io/biocontainers/multiqc:1.11--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/multiqc:1.12--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.12--pyhdfd78af_0' }" input: path multiqc_files diff --git a/nextflow.config b/nextflow.config index e3b67f64..61017da5 100644 --- a/nextflow.config +++ b/nextflow.config @@ -15,9 +15,14 @@ params { // short read preprocessing options clip_tool = 'fastp' - save_trimmed_fail = false + reads_minlength = 15 + fastp_save_trimmed_fail = false fastp_qualified_quality = 15 fastp_cut_mean_quality = 15 + adapterremoval_minquality = 2 + adapterremoval_adapter1 = 'AGATCGGAAGAGCACACGTCTGAACTCCAGTCACNNNNNNATCTCGTATGCCGTCTTCTGCTTG' + adapterremoval_adapter2 = 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT' + adapterremoval_trim_quality_stretch = false keep_phix = false // phix_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Enterobacteria_phage_phiX174_sensu_lato/all_assembly_versions/GCA_002596845.1_ASM259684v1/GCA_002596845.1_ASM259684v1_genomic.fna.gz" phix_reference = "${baseDir}/assets/data/GCA_002596845.1_ASM259684v1_genomic.fna.gz" diff --git a/nextflow_schema.json b/nextflow_schema.json index be926f97..d29f81c7 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -264,16 +264,24 @@ "description": "", "default": "", "properties": { - "save_trimmed_fail": { - "type": "boolean", - "fa_icon": "fas fa-save", - "description": "Save the by fastp trimmed FastQ files in the results directory.", - "help_text": "By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete." + "clip_tool": { + "type": "string", + "default": "fastp", + "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", + "enum": [ + "fastp", + "adapterremoval" + ] + }, + "reads_minlength": { + "type": "integer", + "default": 15, + "description": "The minimum length of reads must have to be retained for downstream analysis." }, "fastp_qualified_quality": { "type": "integer", "default": 15, - "description": "Minimum phred quality value of a base to be qualified.", + "description": "Minimum phred quality value of a base to be qualified in fastp.", "help": "Reads with more than 40% of unqualified bases will be discarded." }, "fastp_cut_mean_quality": { @@ -282,6 +290,30 @@ "description": "The mean quality requirement used for per read sliding window cutting by fastp.", "help": "Used in combination with the fastp options '--cut_front' and '--cut_tail'. If the mean quality within a window (of size 4) is below `--fastp_cut_mean_quality`, the bases are dropped and the sliding window is moved further, otherwise it stops." }, + "fastp_save_trimmed_fail": { + "type": "boolean", + "description": "Save reads that fail fastp filtering in a separate file. Not used downstream." + }, + "adapterremoval_minquality": { + "type": "integer", + "default": 2, + "description": "The minimum base quality for low-quality base trimming by AdapterRemoval." + }, + "adapterremoval_trim_quality_stretch": { + "type": "boolean", + "description": "Turn on quality trimming by consecutive stretch of low quality bases, rather than by window.", + "help_text": "Default base-quality trimming is set to trim by 'windows', as in FastP. Specifying this flag will use trim via contiguous stretch of low quality bases (Ns) instead.\n\n> Replaces --trimwindows 4 with --trimqualities in AdapterRemoval" + }, + "adapterremoval_adapter1": { + "type": "string", + "default": "AGATCGGAAGAGCACACGTCTGAACTCCAGTCACNNNNNNATCTCGTATGCCGTCTTCTGCTTG", + "description": "Forward read adapter to be trimmed by AdapterRemoval." + }, + "adapterremoval_adapter2": { + "type": "string", + "default": "AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT", + "description": "Reverse read adapter to be trimmed by AdapterRemoval for paired end data." + }, "host_genome": { "type": "string", "help_text": "This parameter is mutually exclusive with `--host_genome`. Host read removal is done with Bowtie2. \nBoth the iGenomes FASTA file as well as corresponding, already pre-built Bowtie 2 index files will be used.", @@ -661,4 +693,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} +} \ No newline at end of file diff --git a/workflows/mag.nf b/workflows/mag.nf index 32dbbb17..c27c7384 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -228,7 +228,7 @@ workflow MAG { if ( params.clip_tool == 'fastp' ) { ch_clipmerge_out = FASTP ( ch_raw_short_reads, - [], + params.fastp_save_trimmed_fail, [] ) ch_short_reads = FASTP.out.reads @@ -717,7 +717,7 @@ workflow MAG { ch_quast_multiqc.collect().ifEmpty([]), ch_bowtie2_assembly_multiqc.collect().ifEmpty([]), ch_busco_multiqc.collect().ifEmpty([]), - ch_multiqc_additional + ch_multiqc_additional.collect().ifEmpty([]), ) multiqc_report = MULTIQC.out.report.toList() ch_versions = ch_versions.mix(MULTIQC.out.versions) From cd05e4c987b50c1b924743754b2be1c13f547ee8 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 23 Mar 2022 14:42:23 +0100 Subject: [PATCH 110/246] Add tests --- .github/workflows/ci.yml | 2 +- conf/test_adapterremoval.config | 32 ++++++++++++++++++++++++++++++++ 2 files changed, 33 insertions(+), 1 deletion(-) create mode 100644 conf/test_adapterremoval.config diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 45d45ad8..6bb8bdf8 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -55,7 +55,7 @@ jobs: strategy: matrix: # Run remaining test profiles with minimum nextflow version - profile: [test_host_rm, test_hybrid, test_hybrid_host_rm, test_busco_auto, test_ancient_dna] + profile: [test_host_rm, test_hybrid, test_hybrid_host_rm, test_busco_auto, test_ancient_dna, test_adapterremoval] steps: - name: Check out pipeline code uses: actions/checkout@v2 diff --git a/conf/test_adapterremoval.config b/conf/test_adapterremoval.config new file mode 100644 index 00000000..a9f71433 --- /dev/null +++ b/conf/test_adapterremoval.config @@ -0,0 +1,32 @@ +/* +======================================================================================== + Nextflow config file for running minimal tests +======================================================================================== + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run nf-core/mag -profile test, + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Test profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + + // Limit resources so that this can run on GitHub Actions + max_cpus = 2 + max_memory = '6.GB' + max_time = '6.h' + + // Input data + input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' + centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" + kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" + skip_krona = true + min_length_unbinned_contigs = 1 + max_unbinned_contigs = 2 + busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" + gtdb = false + clip_tool = 'adapterremoval' +} From 8513246a6503762449bf595d74d956bffd7bf1dd Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 23 Mar 2022 14:51:09 +0100 Subject: [PATCH 111/246] Add new test to nextflow.config --- nextflow.config | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 61017da5..2c409bef 100644 --- a/nextflow.config +++ b/nextflow.config @@ -202,7 +202,9 @@ profiles { test_hybrid_host_rm { includeConfig 'conf/test_hybrid_host_rm.config' } test_busco_auto { includeConfig 'conf/test_busco_auto.config' } test_full { includeConfig 'conf/test_full.config' } - test_ancient_dna { includeConfig 'conf/test_ancient_dna.config' } + test_ancient_dna { includeConfig 'conf/test_ancient_dna.config' } + test_adapterremoval { includeConfig 'conf/test_adapterremoval' } + } // Load igenomes.config if required From 7c866e905da3280480c33a84ebff12f7458ccfc3 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Wed, 23 Mar 2022 13:52:17 +0000 Subject: [PATCH 112/246] Template update for nf-core/tools version 2.3.1 --- .editorconfig | 5 +- .github/CONTRIBUTING.md | 15 +- .github/ISSUE_TEMPLATE/bug_report.yml | 1 - .github/PULL_REQUEST_TEMPLATE.md | 6 +- .github/workflows/awsfulltest.yml | 1 - .github/workflows/awstest.yml | 2 +- .github/workflows/branch.yml | 5 +- .github/workflows/ci.yml | 12 +- .github/workflows/linting.yml | 85 ++--------- .github/workflows/linting_comment.yml | 3 +- .gitpod.yml | 16 +- .markdownlint.yml | 14 -- .prettierrc.yml | 1 + .yamllint.yml | 6 - CHANGELOG.md | 2 + CITATIONS.md | 27 ++-- README.md | 31 ++-- assets/email_template.html | 142 ++++++++++++------ assets/multiqc_config.yaml | 11 -- assets/multiqc_config.yml | 11 ++ assets/schema_input.json | 5 +- docs/README.md | 8 +- docs/output.md | 28 ++-- docs/usage.md | 109 +++++++------- modules.json | 8 +- .../custom/dumpsoftwareversions/main.nf | 3 + .../custom/dumpsoftwareversions/meta.yml | 2 +- modules/nf-core/modules/fastqc/main.nf | 3 + modules/nf-core/modules/fastqc/meta.yml | 90 +++++------ modules/nf-core/modules/multiqc/main.nf | 9 +- modules/nf-core/modules/multiqc/meta.yml | 66 ++++---- nextflow_schema.json | 14 +- workflows/mag.nf | 2 +- 33 files changed, 361 insertions(+), 382 deletions(-) delete mode 100644 .markdownlint.yml create mode 100644 .prettierrc.yml delete mode 100644 .yamllint.yml delete mode 100644 assets/multiqc_config.yaml create mode 100644 assets/multiqc_config.yml diff --git a/.editorconfig b/.editorconfig index 95549501..b6b31907 100644 --- a/.editorconfig +++ b/.editorconfig @@ -8,12 +8,9 @@ trim_trailing_whitespace = true indent_size = 4 indent_style = space -[*.{yml,yaml}] +[*.{md,yml,yaml,html,css,scss,js}] indent_size = 2 -[*.json] -insert_final_newline = unset - # These files are edited and tested upstream in nf-core/modules [/modules/nf-core/**] charset = unset diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index be1d64be..3bd0f323 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -15,8 +15,7 @@ Contributions to the code are even more welcome ;) If you'd like to write some code for nf-core/mag, the standard workflow is as follows: -1. Check that there isn't already an issue about your idea in the [nf-core/mag issues](https://github.com/nf-core/mag/issues) to avoid duplicating work - * If there isn't one already, please create one so that others know you're working on this +1. Check that there isn't already an issue about your idea in the [nf-core/mag issues](https://github.com/nf-core/mag/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this 2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/mag repository](https://github.com/nf-core/mag) to your GitHub account 3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions) 4. Use `nf-core schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10). @@ -49,9 +48,9 @@ These tests are run both with the latest available version of `Nextflow` and als :warning: Only in the unlikely and regretful event of a release happening with a bug. -* On your own fork, make a new branch `patch` based on `upstream/master`. -* Fix the bug, and bump version (X.Y.Z+1). -* A PR should be made on `master` from patch to directly this particular bug. +- On your own fork, make a new branch `patch` based on `upstream/master`. +- Fix the bug, and bump version (X.Y.Z+1). +- A PR should be made on `master` from patch to directly this particular bug. ## Getting help @@ -73,7 +72,7 @@ If you wish to contribute a new step, please use the following coding standards: 6. Add sanity checks and validation for all relevant parameters. 7. Perform local tests to validate that the new code works as expected. 8. If applicable, add a new test command in `.github/workflow/ci.yml`. -9. Update MultiQC config `assets/multiqc_config.yaml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module. +9. Update MultiQC config `assets/multiqc_config.yml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module. 10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`. ### Default values @@ -92,8 +91,8 @@ The process resources can be passed on to the tool dynamically within the proces Please use the following naming schemes, to make it easy to understand what is going where. -* initial process channel: `ch_output_from_` -* intermediate and terminal channels: `ch__for_` +- initial process channel: `ch_output_from_` +- intermediate and terminal channels: `ch__for_` ### Nextflow version bumping diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml index a9dff0f9..d32afe01 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.yml +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -2,7 +2,6 @@ name: Bug report description: Report something that is broken or incorrect labels: bug body: - - type: markdown attributes: value: | diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 39b8c44a..1e68020b 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -16,10 +16,10 @@ Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/mag/ - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! - - [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/mag/tree/master/.github/CONTRIBUTING.md) - - [ ] If necessary, also make a PR on the nf-core/mag _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. + - [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/mag/tree/master/.github/CONTRIBUTING.md) + - [ ] If necessary, also make a PR on the nf-core/mag _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core lint`). -- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker` --outdir `). +- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). - [ ] Usage Documentation in `docs/usage.md` is updated. - [ ] Output Documentation in `docs/output.md` is updated. - [ ] `CHANGELOG.md` is updated. diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 0a02db8f..b74d2047 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -18,7 +18,6 @@ jobs: # TODO nf-core: You can customise AWS full pipeline tests as required # Add full size test data (but still relatively small datasets for few samples) # on the `test_full.config` test runs with only one set of parameters - with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 6e009f41..f4aff861 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -10,9 +10,9 @@ jobs: if: github.repository == 'nf-core/mag' runs-on: ubuntu-latest steps: + # Launch workflow using Tower CLI tool action - name: Launch workflow via tower uses: nf-core/tower-action@v3 - with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index d5ee9bae..55f14919 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,8 +13,7 @@ jobs: - name: Check PRs if: github.repository == 'nf-core/mag' run: | - { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/mag ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] - + "{ [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/mag ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]" # If the above check failed, post a comment on the PR explaining the failure # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets @@ -43,4 +42,4 @@ jobs: Thanks again for your contribution! repo-token: ${{ secrets.GITHUB_TOKEN }} allow-repeats: false - +# diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index b1c578c1..f5885cbc 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -16,18 +16,18 @@ jobs: test: name: Run pipeline with test data # Only run on push if this is the nf-core dev branch (merged PRs) - if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/mag') }} + if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/mag') }}" runs-on: ubuntu-latest strategy: matrix: # Nextflow versions include: # Test pipeline minimum Nextflow version - - NXF_VER: '21.10.3' - NXF_EDGE: '' + - NXF_VER: "21.10.3" + NXF_EDGE: "" # Test latest edge release of Nextflow - - NXF_VER: '' - NXF_EDGE: '1' + - NXF_VER: "" + NXF_EDGE: "1" steps: - name: Check out pipeline code uses: actions/checkout@v2 @@ -48,3 +48,5 @@ jobs: # Remember that you can parallelise this by using strategy.matrix run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results + +# diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index fda934c0..e9cf5de3 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -1,6 +1,7 @@ name: nf-core linting # This workflow is triggered on pushes and PRs to the repository. -# It runs the `nf-core lint` and markdown lint tests to ensure that the code meets the nf-core guidelines +# It runs the `nf-core lint` and markdown lint tests to ensure +# that the code meets the nf-core guidelines. on: push: pull_request: @@ -8,42 +9,6 @@ on: types: [published] jobs: - Markdown: - runs-on: ubuntu-latest - steps: - - uses: actions/checkout@v2 - - uses: actions/setup-node@v2 - - name: Install markdownlint - run: npm install -g markdownlint-cli - - name: Run Markdownlint - run: markdownlint . - - # If the above check failed, post a comment on the PR explaining the failure - - name: Post PR comment - if: failure() - uses: mshick/add-pr-comment@v1 - with: - message: | - ## Markdown linting is failing - - To keep the code consistent with lots of contributors, we run automated code consistency checks. - To fix this CI test, please run: - - * Install `markdownlint-cli` - * On Mac: `brew install markdownlint-cli` - * Everything else: [Install `npm`](https://www.npmjs.com/get-npm) then [install `markdownlint-cli`](https://www.npmjs.com/package/markdownlint-cli) (`npm install -g markdownlint-cli`) - * Fix the markdown errors - * Automatically: `markdownlint . --fix` - * Manually resolve anything left from `markdownlint .` - - Once you push these changes the test should pass, and you can hide this comment :+1: - - We highly recommend setting up markdownlint in your code editor so that this formatting is done automatically on save. Ask about it on Slack for help! - - Thanks again for your contribution! - repo-token: ${{ secrets.GITHUB_TOKEN }} - allow-repeats: false - EditorConfig: runs-on: ubuntu-latest steps: @@ -55,49 +20,24 @@ jobs: run: npm install -g editorconfig-checker - name: Run ECLint check - run: editorconfig-checker -exclude README.md $(git ls-files | grep -v test) + run: editorconfig-checker -exclude README.md $(find .* -type f | grep -v '.git\|.py\|.md\|json\|yml\|yaml\|html\|css\|work\|.nextflow\|build\|nf_core.egg-info\|log.txt\|Makefile') - YAML: + Prettier: runs-on: ubuntu-latest steps: - - name: Checkout - uses: actions/checkout@master - - name: 'Yamllint' - uses: karancode/yamllint-github-action@master - with: - yamllint_file_or_dir: '.' - yamllint_config_filepath: '.yamllint.yml' - - # If the above check failed, post a comment on the PR explaining the failure - - name: Post PR comment - if: failure() - uses: mshick/add-pr-comment@v1 - with: - message: | - ## YAML linting is failing - - To keep the code consistent with lots of contributors, we run automated code consistency checks. - To fix this CI test, please run: - - * Install `yamllint` - * Install `yamllint` following [this](https://yamllint.readthedocs.io/en/stable/quickstart.html#installing-yamllint) - instructions or alternative install it in your [conda environment](https://anaconda.org/conda-forge/yamllint) - * Fix the markdown errors - * Run the test locally: `yamllint $(find . -type f -name "*.yml" -o -name "*.yaml") -c ./.yamllint.yml` - * Fix any reported errors in your YAML files + - uses: actions/checkout@v2 - Once you push these changes the test should pass, and you can hide this comment :+1: + - uses: actions/setup-node@v2 - We highly recommend setting up yaml-lint in your code editor so that this formatting is done automatically on save. Ask about it on Slack for help! + - name: Install Prettier + run: npm install -g prettier - Thanks again for your contribution! - repo-token: ${{ secrets.GITHUB_TOKEN }} - allow-repeats: false + - name: Run Prettier --check + run: prettier --check ${GITHUB_WORKSPACE} nf-core: runs-on: ubuntu-latest steps: - - name: Check out pipeline code uses: actions/checkout@v2 @@ -110,8 +50,8 @@ jobs: - uses: actions/setup-python@v1 with: - python-version: '3.6' - architecture: 'x64' + python-version: "3.6" + architecture: "x64" - name: Install dependencies run: | @@ -139,3 +79,4 @@ jobs: lint_results.md PR_number.txt +# diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml index 44d72994..91c487a1 100644 --- a/.github/workflows/linting_comment.yml +++ b/.github/workflows/linting_comment.yml @@ -1,4 +1,3 @@ - name: nf-core linting comment # This workflow is triggered after the linting action is complete # It posts an automated comment to the PR, even if the PR is coming from a fork @@ -27,4 +26,4 @@ jobs: GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} number: ${{ steps.pr_number.outputs.pr_number }} path: linting-logs/lint_results.md - +# diff --git a/.gitpod.yml b/.gitpod.yml index b7d4cee1..c452ee93 100644 --- a/.gitpod.yml +++ b/.gitpod.yml @@ -2,13 +2,13 @@ image: nfcore/gitpod:latest vscode: extensions: # based on nf-core.nf-core-extensionpack - - codezombiech.gitignore # Language support for .gitignore files + - codezombiech.gitignore # Language support for .gitignore files # - cssho.vscode-svgviewer # SVG viewer - - davidanson.vscode-markdownlint # Markdown/CommonMark linting and style checking for Visual Studio Code - - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed - - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files - - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar - - mechatroner.rainbow-csv # Highlight columns in csv files in different colors + - davidanson.vscode-markdownlint # Markdown/CommonMark linting and style checking for Visual Studio Code + - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed + - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files + - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar + - mechatroner.rainbow-csv # Highlight columns in csv files in different colors # - nextflow.nextflow # Nextflow syntax highlighting - - oderwat.indent-rainbow # Highlight indentation level - - streetsidesoftware.code-spell-checker # Spelling checker for source code + - oderwat.indent-rainbow # Highlight indentation level + - streetsidesoftware.code-spell-checker # Spelling checker for source code diff --git a/.markdownlint.yml b/.markdownlint.yml deleted file mode 100644 index 9e605fcf..00000000 --- a/.markdownlint.yml +++ /dev/null @@ -1,14 +0,0 @@ -# Markdownlint configuration file -default: true -line-length: false -ul-indent: - indent: 4 -no-duplicate-header: - siblings_only: true -no-inline-html: - allowed_elements: - - img - - p - - kbd - - details - - summary diff --git a/.prettierrc.yml b/.prettierrc.yml new file mode 100644 index 00000000..c81f9a76 --- /dev/null +++ b/.prettierrc.yml @@ -0,0 +1 @@ +printWidth: 120 diff --git a/.yamllint.yml b/.yamllint.yml deleted file mode 100644 index d466deec..00000000 --- a/.yamllint.yml +++ /dev/null @@ -1,6 +0,0 @@ -extends: default - -rules: - document-start: disable - line-length: disable - truthy: disable diff --git a/CHANGELOG.md b/CHANGELOG.md index 0803027f..0f46b560 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,6 +11,8 @@ Initial release of nf-core/mag, created with the [nf-core](https://nf-co.re/) te ### `Fixed` +- Clarified conda usage and added an installation tutorial for Singularity since the one on Syllabs' website uses an outdate version of GO Compiler + ### `Dependencies` ### `Deprecated` diff --git a/CITATIONS.md b/CITATIONS.md index 256b0739..f6921e00 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,23 +10,26 @@ ## Pipeline tools -* [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) +- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) -* [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) - > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. +- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) + > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. ## Software packaging/containerisation tools -* [Anaconda](https://anaconda.com) - > Anaconda Software Distribution. Computer software. Vers. 2-2.4.0. Anaconda, Nov. 2016. Web. +- [Anaconda](https://anaconda.com) -* [Bioconda](https://pubmed.ncbi.nlm.nih.gov/29967506/) - > Grüning B, Dale R, Sjödin A, Chapman BA, Rowe J, Tomkins-Tinch CH, Valieris R, Köster J; Bioconda Team. Bioconda: sustainable and comprehensive software distribution for the life sciences. Nat Methods. 2018 Jul;15(7):475-476. doi: 10.1038/s41592-018-0046-7. PubMed PMID: 29967506. + > Anaconda Software Distribution. Computer software. Vers. 2-2.4.0. Anaconda, Nov. 2016. Web. -* [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/) - > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671. +- [Bioconda](https://pubmed.ncbi.nlm.nih.gov/29967506/) -* [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241) + > Grüning B, Dale R, Sjödin A, Chapman BA, Rowe J, Tomkins-Tinch CH, Valieris R, Köster J; Bioconda Team. Bioconda: sustainable and comprehensive software distribution for the life sciences. Nat Methods. 2018 Jul;15(7):475-476. doi: 10.1038/s41592-018-0046-7. PubMed PMID: 29967506. -* [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/) - > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675. +- [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/) + + > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671. + +- [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241) + +- [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/) + > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675. diff --git a/README.md b/README.md index 39f3d98e..8819a575 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,4 @@ -# ![nf-core/mag](docs/images/nf-core-mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core-mag_logo_dark.png#gh-dark-mode-only) +# ![nf-core/mag](docs/images/nf-core/mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core/mag_logo_dark.png#gh-dark-mode-only) [![GitHub Actions CI Status](https://github.com/nf-core/mag/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) @@ -17,11 +17,13 @@ ## Introduction + **nf-core/mag** is a bioinformatics best-practice analysis pipeline for Assembly, binning and annotation of metagenomes. The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from [nf-core/modules](https://github.com/nf-core/modules) in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community! + On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources. The results obtained from the full-sized test can be viewed on the [nf-core website](https://nf-co.re/mag/results). ## Pipeline summary @@ -35,28 +37,28 @@ On release, automated continuous integration tests run the pipeline on a full-si 1. Install [`Nextflow`](https://www.nextflow.io/docs/latest/getstarted.html#installation) (`>=21.10.3`) -2. Install any of [`Docker`](https://docs.docker.com/engine/installation/), [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/), [`Podman`](https://podman.io/), [`Shifter`](https://nersc.gitlab.io/development/shifter/how-to-use/) or [`Charliecloud`](https://hpc.github.io/charliecloud/) for full pipeline reproducibility _(please only use [`Conda`](https://conda.io/miniconda.html) as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_ +2. Install any of [`Docker`](https://docs.docker.com/engine/installation/), [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) (you can follow [this tutorial](https://singularity-tutorial.github.io/01-installation/)), [`Podman`](https://podman.io/), [`Shifter`](https://nersc.gitlab.io/development/shifter/how-to-use/) or [`Charliecloud`](https://hpc.github.io/charliecloud/) for full pipeline reproducibility _(you can use [`Conda`](https://conda.io/miniconda.html) both to install Nextflow itself and also to manage software within pipelines. Please only use it within pipelines as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_. 3. Download the pipeline and test it on a minimal dataset with a single command: - ```console - nextflow run nf-core/mag -profile test,YOURPROFILE --outdir - ``` + ```console + nextflow run nf-core/mag -profile test,YOURPROFILE --outdir + ``` - Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. + Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. - > * The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. - > * Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. - > * If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. - > * If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. + > - The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. + > - Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. + > - If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. + > - If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. 4. Start running your own analysis! - + - ```console - nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile - ``` + ```console + nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile + ``` ## Documentation @@ -82,6 +84,7 @@ For further information or help, don't hesitate to get in touch on the [Slack `# + An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. You can cite the `nf-core` publication as follows: diff --git a/assets/email_template.html b/assets/email_template.html index 13cdbf26..1563166a 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -1,53 +1,111 @@ - - - - + + + + - - nf-core/mag Pipeline Report - - - + diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml deleted file mode 100644 index 7b6b9073..00000000 --- a/assets/multiqc_config.yaml +++ /dev/null @@ -1,11 +0,0 @@ -report_comment: > - This report has been generated by the nf-core/mag - analysis pipeline. For information about how to interpret these results, please see the - documentation. -report_section_order: - software_versions: - order: -1000 - nf-core-mag-summary: - order: -1001 - -export_plots: true diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml new file mode 100644 index 00000000..db2c548c --- /dev/null +++ b/assets/multiqc_config.yml @@ -0,0 +1,11 @@ +report_comment: > + This report has been generated by the nf-core/mag + analysis pipeline. For information about how to interpret these results, please see the + documentation. +report_section_order: + software_versions: + order: -1000 + "nf-core-mag-summary": + order: -1001 + +export_plots: true diff --git a/assets/schema_input.json b/assets/schema_input.json index 0091ae04..44d6d151 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -31,9 +31,6 @@ ] } }, - "required": [ - "sample", - "fastq_1" - ] + "required": ["sample", "fastq_1"] } } diff --git a/docs/README.md b/docs/README.md index 7b0a1c80..b1cbb97f 100644 --- a/docs/README.md +++ b/docs/README.md @@ -2,9 +2,9 @@ The nf-core/mag documentation is split into the following pages: -* [Usage](usage.md) - * An overview of how the pipeline works, how to run it and a description of all of the different command-line flags. -* [Output](output.md) - * An overview of the different results produced by the pipeline and how to interpret them. +- [Usage](usage.md) + - An overview of how the pipeline works, how to run it and a description of all of the different command-line flags. +- [Output](output.md) + - An overview of the different results produced by the pipeline and how to interpret them. You can find a lot more documentation about installing, configuring and running nf-core pipelines on the website: [https://nf-co.re](https://nf-co.re) diff --git a/docs/output.md b/docs/output.md index a2782c16..9efc681c 100644 --- a/docs/output.md +++ b/docs/output.md @@ -12,18 +12,18 @@ The directories listed below will be created in the results directory after the The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: -* [FastQC](#fastqc) - Raw read QC -* [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline -* [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution +- [FastQC](#fastqc) - Raw read QC +- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline +- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution ### FastQC
Output files -* `fastqc/` - * `*_fastqc.html`: FastQC report containing quality metrics. - * `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images. +- `fastqc/` + - `*_fastqc.html`: FastQC report containing quality metrics. + - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images.
@@ -42,10 +42,10 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d
Output files -* `multiqc/` - * `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser. - * `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline. - * `multiqc_plots/`: directory containing static images from the report in various formats. +- `multiqc/` + - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser. + - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline. + - `multiqc_plots/`: directory containing static images from the report in various formats.
@@ -58,10 +58,10 @@ Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQ
Output files -* `pipeline_info/` - * Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. - * Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. - * Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`. +- `pipeline_info/` + - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. + - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. + - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`.
diff --git a/docs/usage.md b/docs/usage.md index 0cb8bd3f..81b33493 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -44,11 +44,11 @@ TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz, TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz, ``` -| Column | Description | -|----------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| -| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | -| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | -| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | +| Column | Description | +| --------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | +| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | +| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline. @@ -57,7 +57,7 @@ An [example samplesheet](../assets/samplesheet.csv) has been provided with the p The typical command for running the pipeline is as follows: ```console -nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile docker +nextflow run nf-core/mag --input samplesheet.csv --outdir --genome GRCh37 -profile docker ``` This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. @@ -65,9 +65,9 @@ This will launch the pipeline with the `docker` configuration profile. See below Note that the pipeline will create the following files in your working directory: ```console -work # Directory containing the nextflow working files -results # Finished results (configurable, see below) -.nextflow_log # Log file from Nextflow +work # Directory containing the nextflow working files + # Finished results in specified location (defined with --outdir) +.nextflow_log # Log file from Nextflow # Other nextflow hidden files, eg. history of pipeline runs and old logs. ``` @@ -106,25 +106,25 @@ They are loaded in sequence, so later profiles can overwrite earlier profiles. If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended. -* `docker` - * A generic configuration profile to be used with [Docker](https://docker.com/) -* `singularity` - * A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/) -* `podman` - * A generic configuration profile to be used with [Podman](https://podman.io/) -* `shifter` - * A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) -* `charliecloud` - * A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) -* `conda` - * A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud. -* `test` - * A profile with a complete configuration for automated testing - * Includes links to test data so needs no other parameters +- `docker` + - A generic configuration profile to be used with [Docker](https://docker.com/) +- `singularity` + - A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/) +- `podman` + - A generic configuration profile to be used with [Podman](https://podman.io/) +- `shifter` + - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) +- `charliecloud` + - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) +- `conda` + - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud. +- `test` + - A profile with a complete configuration for automated testing + - Includes links to test data so needs no other parameters ### `-resume` -Specify this when restarting a pipeline. Nextflow will used cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. +Specify this when restarting a pipeline. Nextflow will use cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. For input to be considered the same, not only the names must be identical but the files' contents as well. For more info about this parameter, see [this blog post](https://www.nextflow.io/blog/2019/demystifying-nextflow-resume.html). You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names. @@ -186,6 +186,7 @@ process { ``` > **NB:** We specify the full process name i.e. `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN` in the config file because this takes priority over the short name (`STAR_ALIGN`) and allows existing configuration using the full process name to be correctly overridden. +> > If you get a warning suggesting that the process selector isn't recognised check that the process name has been specified correctly. ### Updating containers @@ -196,35 +197,35 @@ The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementatio 2. Find the latest version of the Biocontainer available on [Quay.io](https://quay.io/repository/biocontainers/pangolin?tag=latest&tab=tags) 3. Create the custom config accordingly: - * For Docker: - - ```nextflow - process { - withName: PANGOLIN { - container = 'quay.io/biocontainers/pangolin:3.0.5--pyhdfd78af_0' - } - } - ``` - - * For Singularity: - - ```nextflow - process { - withName: PANGOLIN { - container = 'https://depot.galaxyproject.org/singularity/pangolin:3.0.5--pyhdfd78af_0' - } - } - ``` - - * For Conda: - - ```nextflow - process { - withName: PANGOLIN { - conda = 'bioconda::pangolin=3.0.5' - } - } - ``` + - For Docker: + + ```nextflow + process { + withName: PANGOLIN { + container = 'quay.io/biocontainers/pangolin:3.0.5--pyhdfd78af_0' + } + } + ``` + + - For Singularity: + + ```nextflow + process { + withName: PANGOLIN { + container = 'https://depot.galaxyproject.org/singularity/pangolin:3.0.5--pyhdfd78af_0' + } + } + ``` + + - For Conda: + + ```nextflow + process { + withName: PANGOLIN { + conda = 'bioconda::pangolin=3.0.5' + } + } + ``` > **NB:** If you wish to periodically update individual tool-specific results (e.g. Pangolin) generated by the pipeline then you must ensure to keep the `work/` directory otherwise the `-resume` ability of the pipeline will be compromised and it will restart from scratch. diff --git a/modules.json b/modules.json index 6394e068..1768fed6 100644 --- a/modules.json +++ b/modules.json @@ -4,14 +4,14 @@ "repos": { "nf-core/modules": { "custom/dumpsoftwareversions": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "fastqc": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "multiqc": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" } } } -} \ No newline at end of file +} diff --git a/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf b/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf index 934bb467..327d5100 100644 --- a/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf +++ b/modules/nf-core/modules/custom/dumpsoftwareversions/main.nf @@ -15,6 +15,9 @@ process CUSTOM_DUMPSOFTWAREVERSIONS { path "software_versions_mqc.yml", emit: mqc_yml path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' template 'dumpsoftwareversions.py' diff --git a/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml index 5b5b8a60..60b546a0 100644 --- a/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml +++ b/modules/nf-core/modules/custom/dumpsoftwareversions/meta.yml @@ -8,7 +8,7 @@ tools: description: Custom module used to dump software versions within the nf-core pipeline template homepage: https://github.com/nf-core/tools documentation: https://github.com/nf-core/tools - licence: ['MIT'] + licence: ["MIT"] input: - versions: type: file diff --git a/modules/nf-core/modules/fastqc/main.nf b/modules/nf-core/modules/fastqc/main.nf index d250eca0..ed6b8c50 100644 --- a/modules/nf-core/modules/fastqc/main.nf +++ b/modules/nf-core/modules/fastqc/main.nf @@ -15,6 +15,9 @@ process FASTQC { tuple val(meta), path("*.zip") , emit: zip path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' // Add soft-links to original FastQs for consistent naming in pipeline diff --git a/modules/nf-core/modules/fastqc/meta.yml b/modules/nf-core/modules/fastqc/meta.yml index b09553a3..4da5bb5a 100644 --- a/modules/nf-core/modules/fastqc/meta.yml +++ b/modules/nf-core/modules/fastqc/meta.yml @@ -1,52 +1,52 @@ name: fastqc description: Run FastQC on sequenced reads keywords: - - quality control - - qc - - adapters - - fastq + - quality control + - qc + - adapters + - fastq tools: - - fastqc: - description: | - FastQC gives general quality metrics about your reads. - It provides information about the quality score distribution - across your reads, the per base sequence content (%A/C/G/T). - You get information about adapter contamination and other - overrepresented sequences. - homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ - documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ - licence: ['GPL-2.0-only'] + - fastqc: + description: | + FastQC gives general quality metrics about your reads. + It provides information about the quality score distribution + across your reads, the per base sequence content (%A/C/G/T). + You get information about adapter contamination and other + overrepresented sequences. + homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ + documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ + licence: ["GPL-2.0-only"] input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - reads: - type: file - description: | - List of input FastQ files of size 1 and 2 for single-end and paired-end data, - respectively. + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - html: - type: file - description: FastQC report - pattern: "*_{fastqc.html}" - - zip: - type: file - description: FastQC report archive - pattern: "*_{fastqc.zip}" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - html: + type: file + description: FastQC report + pattern: "*_{fastqc.html}" + - zip: + type: file + description: FastQC report archive + pattern: "*_{fastqc.zip}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@drpatelh" - - "@grst" - - "@ewels" - - "@FelixKrueger" + - "@drpatelh" + - "@grst" + - "@ewels" + - "@FelixKrueger" diff --git a/modules/nf-core/modules/multiqc/main.nf b/modules/nf-core/modules/multiqc/main.nf index 3dceb162..1264aac1 100644 --- a/modules/nf-core/modules/multiqc/main.nf +++ b/modules/nf-core/modules/multiqc/main.nf @@ -1,10 +1,10 @@ process MULTIQC { label 'process_medium' - conda (params.enable_conda ? 'bioconda::multiqc=1.11' : null) + conda (params.enable_conda ? 'bioconda::multiqc=1.12' : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/multiqc:1.11--pyhdfd78af_0' : - 'quay.io/biocontainers/multiqc:1.11--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/multiqc:1.12--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.12--pyhdfd78af_0' }" input: path multiqc_files @@ -15,6 +15,9 @@ process MULTIQC { path "*_plots" , optional:true, emit: plots path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' """ diff --git a/modules/nf-core/modules/multiqc/meta.yml b/modules/nf-core/modules/multiqc/meta.yml index 63c75a45..6fa891ef 100644 --- a/modules/nf-core/modules/multiqc/meta.yml +++ b/modules/nf-core/modules/multiqc/meta.yml @@ -1,40 +1,40 @@ name: MultiQC description: Aggregate results from bioinformatics analyses across many samples into a single report keywords: - - QC - - bioinformatics tools - - Beautiful stand-alone HTML report + - QC + - bioinformatics tools + - Beautiful stand-alone HTML report tools: - - multiqc: - description: | - MultiQC searches a given directory for analysis logs and compiles a HTML report. - It's a general use tool, perfect for summarising the output from numerous bioinformatics tools. - homepage: https://multiqc.info/ - documentation: https://multiqc.info/docs/ - licence: ['GPL-3.0-or-later'] + - multiqc: + description: | + MultiQC searches a given directory for analysis logs and compiles a HTML report. + It's a general use tool, perfect for summarising the output from numerous bioinformatics tools. + homepage: https://multiqc.info/ + documentation: https://multiqc.info/docs/ + licence: ["GPL-3.0-or-later"] input: - - multiqc_files: - type: file - description: | - List of reports / files recognised by MultiQC, for example the html and zip output of FastQC + - multiqc_files: + type: file + description: | + List of reports / files recognised by MultiQC, for example the html and zip output of FastQC output: - - report: - type: file - description: MultiQC report file - pattern: "multiqc_report.html" - - data: - type: dir - description: MultiQC data dir - pattern: "multiqc_data" - - plots: - type: file - description: Plots created by MultiQC - pattern: "*_data" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" + - report: + type: file + description: MultiQC report file + pattern: "multiqc_report.html" + - data: + type: dir + description: MultiQC data dir + pattern: "multiqc_data" + - plots: + type: file + description: Plots created by MultiQC + pattern: "*_data" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" authors: - - "@abhi18av" - - "@bunop" - - "@drpatelh" + - "@abhi18av" + - "@bunop" + - "@drpatelh" diff --git a/nextflow_schema.json b/nextflow_schema.json index bc78304a..2721a11e 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,10 +10,7 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": [ - "input", - "outdir" - ], + "required": ["input", "outdir"], "properties": { "input": { "type": "string", @@ -185,14 +182,7 @@ "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", - "enum": [ - "symlink", - "rellink", - "link", - "copy", - "copyNoFollow", - "move" - ], + "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], "hidden": true }, "email_on_fail": { diff --git a/workflows/mag.nf b/workflows/mag.nf index 9dc9e53a..9a755509 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -23,7 +23,7 @@ if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input sample ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -ch_multiqc_config = file("$projectDir/assets/multiqc_config.yaml", checkIfExists: true) +ch_multiqc_config = file("$projectDir/assets/multiqc_config.yml", checkIfExists: true) ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multiqc_config) : Channel.empty() /* From 222689f6053381872c1d4103a8ae24eff90ee076 Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 23 Mar 2022 14:53:34 +0100 Subject: [PATCH 113/246] add publication --- CHANGELOG.md | 1 + README.md | 4 ++-- lib/WorkflowMain.groovy | 4 ++-- 3 files changed, 5 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 96f94b7e..af021b0b 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Changed` - [#269](https://github.com/nf-core/mag/pull/269) - Update to nf-core 2.2 `TEMPLATE` +- [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint ### `Fixed` diff --git a/README.md b/README.md index b86fb617..3ecec220 100644 --- a/README.md +++ b/README.md @@ -4,7 +4,7 @@ [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) [![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/mag/results) [![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.3589527-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.3589527) -[![Cite Preprint](https://img.shields.io/badge/Cite%20Us!-Cite%20Preprint-orange)](https://doi.org/10.1101/2021.08.29.458094) +[![Cite Publucation](https://img.shields.io/badge/Cite%20Us!-Cite%20Preprint-orange)](https://doi.org/10.1093/nargab/lqac007) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.10.3-23aa62.svg?labelColor=000000)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) @@ -116,7 +116,7 @@ If you use nf-core/mag for your analysis, please cite the preprint as follows: > > Sabrina Krakau, Daniel Straub, Hadrien Gourlé, Gisela Gabernet, Sven Nahnsen. > -> bioRxiv 2021.08.29.458094. doi: [10.1101/2021.08.29.458094](https://doi.org/10.1101/2021.08.29.458094). +> NAR Genom Bioinform. 2022 Feb 2;4(1):lqac007. doi: [10.1093/nargab/lqac007](https://doi.org/10.1093/nargab/lqac007). additionally you can cite the pipeline directly with the following doi: [10.5281/zenodo.3589527](https://doi.org/10.5281/zenodo.3589527) diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index d9883bce..e85e510c 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -9,8 +9,8 @@ class WorkflowMain { // public static String citation(workflow) { return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" + - "* The preprint\n" + - " https://doi.org/10.1101/2021.08.29.458094\n\n" + + "* The pipeline publication\n" + + " https://doi.org/10.1093/nargab/lqac007\n\n" + "* The pipeline\n" + " https://doi.org/10.5281/zenodo.3589527\n\n" + "* The nf-core framework\n" + From 0a6c4a0cba20dd4deafa24fcc5247894b2fb3d44 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 23 Mar 2022 14:58:39 +0100 Subject: [PATCH 114/246] Forgot a file suffix --- conf/test_adapterremoval.config | 4 ++-- nextflow.config | 2 +- 2 files changed, 3 insertions(+), 3 deletions(-) diff --git a/conf/test_adapterremoval.config b/conf/test_adapterremoval.config index a9f71433..0725cbc2 100644 --- a/conf/test_adapterremoval.config +++ b/conf/test_adapterremoval.config @@ -11,8 +11,8 @@ */ params { - config_profile_name = 'Test profile' - config_profile_description = 'Minimal test dataset to check pipeline function' + config_profile_name = 'Test profile for running with AdapterRemoval' + config_profile_description = 'Minimal test dataset to check pipeline function with AdapterRemoval data' // Limit resources so that this can run on GitHub Actions max_cpus = 2 diff --git a/nextflow.config b/nextflow.config index 2c409bef..eed41039 100644 --- a/nextflow.config +++ b/nextflow.config @@ -203,7 +203,7 @@ profiles { test_busco_auto { includeConfig 'conf/test_busco_auto.config' } test_full { includeConfig 'conf/test_full.config' } test_ancient_dna { includeConfig 'conf/test_ancient_dna.config' } - test_adapterremoval { includeConfig 'conf/test_adapterremoval' } + test_adapterremoval { includeConfig 'conf/test_adapterremoval.config' } } From 80d9026e1bd521513956b528c1cfe8f48f32b66d Mon Sep 17 00:00:00 2001 From: Daniel Straub <42973691+d4straub@users.noreply.github.com> Date: Wed, 23 Mar 2022 15:02:27 +0100 Subject: [PATCH 115/246] Update README.md Co-authored-by: James A. Fellows Yates --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 3ecec220..1c705613 100644 --- a/README.md +++ b/README.md @@ -4,7 +4,7 @@ [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) [![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/mag/results) [![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.3589527-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.3589527) -[![Cite Publucation](https://img.shields.io/badge/Cite%20Us!-Cite%20Preprint-orange)](https://doi.org/10.1093/nargab/lqac007) +[![Cite Publication](https://img.shields.io/badge/Cite%20Us!-Cite%20Publication-orange)](https://doi.org/10.1093/nargab/lqac007) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.10.3-23aa62.svg?labelColor=000000)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) From 2a45dc407ffb768caca642a8ade4a4d5cbabb9dc Mon Sep 17 00:00:00 2001 From: d4straub Date: Wed, 23 Mar 2022 15:56:07 +0100 Subject: [PATCH 116/246] fix EditorConfig --- conf/test_ancient_dna.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config index e6b30534..d4c621ef 100644 --- a/conf/test_ancient_dna.config +++ b/conf/test_ancient_dna.config @@ -19,7 +19,7 @@ params { max_memory = '6.GB' max_time = '6.h' - // Input data + // Input data input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" From 14d01bd30a5af96f1fa567284c2b111f714515dd Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:13:37 +0100 Subject: [PATCH 117/246] prettier and modules update --- .github/workflows/ci.yml | 2 +- .nf-core.yml | 7 +- CHANGELOG.md | 8 +- CITATIONS.md | 114 +++--- README.md | 44 +-- assets/multiqc_config.yml | 67 ++-- docs/output.md | 338 +++++++++--------- docs/usage.md | 62 ++-- modules.json | 20 +- .../nf-core/modules/bcftools/index/main.nf | 13 +- .../nf-core/modules/bcftools/index/meta.yml | 2 +- modules/nf-core/modules/fastp/main.nf | 5 +- modules/nf-core/modules/fastp/meta.yml | 8 +- modules/nf-core/modules/freebayes/main.nf | 8 +- modules/nf-core/modules/freebayes/meta.yml | 20 +- .../jgisummarizebamcontigdepths/main.nf | 3 + .../jgisummarizebamcontigdepths/meta.yml | 2 +- .../nf-core/modules/metabat2/metabat2/main.nf | 3 + .../modules/metabat2/metabat2/meta.yml | 7 +- modules/nf-core/modules/prodigal/main.nf | 3 + modules/nf-core/modules/prodigal/meta.yml | 6 +- modules/nf-core/modules/prokka/main.nf | 3 + modules/nf-core/modules/prokka/meta.yml | 2 +- .../nf-core/modules/pydamage/analyze/main.nf | 5 +- .../nf-core/modules/pydamage/analyze/meta.yml | 2 +- .../nf-core/modules/pydamage/filter/main.nf | 5 +- .../nf-core/modules/pydamage/filter/meta.yml | 2 +- 27 files changed, 404 insertions(+), 357 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 8a3ae18c..3ca68a72 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -46,7 +46,6 @@ jobs: run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results - profiles: name: Run workflow profile # Only run on push if this is the nf-core dev branch (merged PRs) @@ -68,4 +67,5 @@ jobs: - name: Run pipeline with ${{ matrix.profile }} test profile run: | nextflow run ${GITHUB_WORKSPACE} -profile ${{ matrix.profile }},docker --outdir ./results + # diff --git a/.nf-core.yml b/.nf-core.yml index 046e58f7..feabf6ad 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -1,8 +1,5 @@ repository_type: pipeline lint: - files_unchanged: - - lib/NfcoreTemplate.groovy - - .github/workflows/branch.yml - - .github/workflows/linting_comment.yml - - .github/workflows/linting.yml + files_unchanged: + - lib/NfcoreTemplate.groovy diff --git a/CHANGELOG.md b/CHANGELOG.md index 31ac02c4..14fa4c95 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -22,10 +22,10 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Dependencies` -| Tool | Previous version | New version | -| --- | --- | --- | -| bcftools | 1.13 | 1.14 | -| samtools | 1.14 | 1.15 | +| Tool | Previous version | New version | +| -------- | ---------------- | ----------- | +| bcftools | 1.13 | 1.14 | +| samtools | 1.14 | 1.15 | ## v2.1.1 - 2021/11/25 diff --git a/CITATIONS.md b/CITATIONS.md index 71e542eb..387ad6c3 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,79 +10,99 @@ ## Pipeline tools -* [BCFtools](https://doi.org/10.1093/gigascience/giab008) - > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 +- [BCFtools](https://doi.org/10.1093/gigascience/giab008) -* [Bowtie2](https:/dx.doi.org/10.1038/nmeth.1923) - > Langmead, B. and Salzberg, S. L. 2012 Fast gapped-read alignment with Bowtie 2. Nature methods, 9(4), p. 357–359. doi: 10.1038/nmeth.1923. + > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 -* [Busco](https://doi.org/10.1007/978-1-4939-9173-0_14) - > Seppey, M., Manni, M., & Zdobnov, E. M. (2019). BUSCO: assessing genome assembly and annotation completeness. In Gene prediction (pp. 227-245). Humana, New York, NY. doi: 10.1007/978-1-4939-9173-0_14. +- [Bowtie2](https:/dx.doi.org/10.1038/nmeth.1923) -* [CAT](https://doi.org/10.1186/s13059-019-1817-x) - > von Meijenfeldt, F. B., Arkhipova, K., Cambuy, D. D., Coutinho, F. H., & Dutilh, B. E. (2019). Robust taxonomic classification of uncharted microbial sequences and bins with CAT and BAT. Genome biology, 20(1), 1-14. doi: 10.1186/s13059-019-1817-x. + > Langmead, B. and Salzberg, S. L. 2012 Fast gapped-read alignment with Bowtie 2. Nature methods, 9(4), p. 357–359. doi: 10.1038/nmeth.1923. -* [Centrifuge](https://doi.org/10.1101/gr.210641.116) - > Kim, D., Song, L., Breitwieser, F. P., & Salzberg, S. L. (2016). Centrifuge: rapid and sensitive classification of metagenomic sequences. Genome research, 26(12), 1721-1729. doi: 10.1101/gr.210641.116. +- [Busco](https://doi.org/10.1007/978-1-4939-9173-0_14) -* [FastP](https://doi.org/10.1093/bioinformatics/bty560) - > Chen, S., Zhou, Y., Chen, Y., & Gu, J. (2018). fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics , 34(17), i884–i890. doi: 10.1093/bioinformatics/bty560. + > Seppey, M., Manni, M., & Zdobnov, E. M. (2019). BUSCO: assessing genome assembly and annotation completeness. In Gene prediction (pp. 227-245). Humana, New York, NY. doi: 10.1007/978-1-4939-9173-0_14. -* [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) +- [CAT](https://doi.org/10.1186/s13059-019-1817-x) -* [Filtlong](https://github.com/rrwick/Filtlong) + > von Meijenfeldt, F. B., Arkhipova, K., Cambuy, D. D., Coutinho, F. H., & Dutilh, B. E. (2019). Robust taxonomic classification of uncharted microbial sequences and bins with CAT and BAT. Genome biology, 20(1), 1-14. doi: 10.1186/s13059-019-1817-x. -* [Freebayes](https://arxiv.org/abs/1207.3907) - > Garrison E, Marth G. Haplotype-based variant detection from short-read sequencing. arXiv preprint arXiv:1207.3907 [q-bio.GN] 2012 +- [Centrifuge](https://doi.org/10.1101/gr.210641.116) -* [GTDB-Tk](https://doi.org/10.1093/bioinformatics/btz848) - > Chaumeil, P. A., Mussig, A. J., Hugenholtz, P., & Parks, D. H. (2020). GTDB-Tk: a toolkit to classify genomes with the Genome Taxonomy Database. Bioinformatics , 36(6), 1925–1927. doi: 10.1093/bioinformatics/btz848. + > Kim, D., Song, L., Breitwieser, F. P., & Salzberg, S. L. (2016). Centrifuge: rapid and sensitive classification of metagenomic sequences. Genome research, 26(12), 1721-1729. doi: 10.1101/gr.210641.116. -* [Kraken2](https://doi.org/10.1186/s13059-019-1891-0) - > Wood, D et al., 2019. Improved metagenomic analysis with Kraken 2. Genome Biology volume 20, Article number: 257. doi: 10.1186/s13059-019-1891-0. +- [FastP](https://doi.org/10.1093/bioinformatics/bty560) -* [Krona](https://doi.org/10.1186/1471-2105-12-385) - > Ondov, B. D., Bergman, N. H., & Phillippy, A. M. (2011). Interactive metagenomic visualization in a Web browser. BMC bioinformatics, 12(1), 1-10. doi: 10.1186/1471-2105-12-385. + > Chen, S., Zhou, Y., Chen, Y., & Gu, J. (2018). fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics , 34(17), i884–i890. doi: 10.1093/bioinformatics/bty560. -* [MaxBin2](https://doi.org/10.1093/bioinformatics/btv638) - > Yu-Wei, W., Simmons, B. A. & Singer, S. W. (2015) MaxBin 2.0: An Automated Binning Algorithm to Recover Genomes from Multiple Metagenomic Datasets. Bioinformatics 32 (4): 605–7. doi: 10.1093/bioinformatics/btv638. +- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) -* [MEGAHIT](https://doi.org/10.1016/j.ymeth.2016.02.020) - > Li, D., Luo, R., Liu, C. M., Leung, C. M., Ting, H. F., Sadakane, K., ... & Lam, T. W. (2016). MEGAHIT v1. 0: a fast and scalable metagenome assembler driven by advanced methodologies and community practices. Methods, 102, 3-11. doi: 10.1016/j.ymeth.2016.02.020. +- [Filtlong](https://github.com/rrwick/Filtlong) -* [MetaBAT2](https://doi.org/10.7717/peerj.7359) - > Kang, D. D., Li, F., Kirton, E., Thomas, A., Egan, R., An, H., & Wang, Z. (2019). MetaBAT 2: an adaptive binning algorithm for robust and efficient genome reconstruction from metagenome assemblies. PeerJ, 7, e7359. doi: 10.7717/peerj.7359. +- [Freebayes](https://arxiv.org/abs/1207.3907) -* [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) - > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. + > Garrison E, Marth G. Haplotype-based variant detection from short-read sequencing. arXiv preprint arXiv:1207.3907 [q-bio.GN] 2012 -* [NanoLyse](https://doi.org/10.1093/bioinformatics/bty149) - > De Coster, W., D’Hert, S., Schultz, D. T., Cruts, M., & Van Broeckhoven, C. (2018). NanoPack: visualizing and processing long-read sequencing data. Bioinformatics, 34(15), 2666-2669. doi: 10.1093/bioinformatics/bty149. +- [GTDB-Tk](https://doi.org/10.1093/bioinformatics/btz848) -* [NanoPlot](https://doi.org/10.1093/bioinformatics/bty149) - > De Coster, W., D’Hert, S., Schultz, D. T., Cruts, M., & Van Broeckhoven, C. (2018). NanoPack: visualizing and processing long-read sequencing data. Bioinformatics, 34(15), 2666-2669. doi: 10.1093/bioinformatics/bty149. + > Chaumeil, P. A., Mussig, A. J., Hugenholtz, P., & Parks, D. H. (2020). GTDB-Tk: a toolkit to classify genomes with the Genome Taxonomy Database. Bioinformatics , 36(6), 1925–1927. doi: 10.1093/bioinformatics/btz848. -* [Porechop](https://github.com/rrwick/Porechop) +- [Kraken2](https://doi.org/10.1186/s13059-019-1891-0) -* [Prodigal](https://pubmed.ncbi.nlm.nih.gov/20211023/) - > Hyatt D, Chen GL, Locascio PF, Land ML, Larimer FW, Hauser LJ. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinformatics. 2010 Mar 8;11:119. doi: 10.1186/1471-2105-11-119. PMID: 20211023; PMCID: PMC2848648. + > Wood, D et al., 2019. Improved metagenomic analysis with Kraken 2. Genome Biology volume 20, Article number: 257. doi: 10.1186/s13059-019-1891-0. -* [Prokka](https://pubmed.ncbi.nlm.nih.gov/24642063/) - > Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014 Jul 15;30(14):2068-9. doi: 10.1093/bioinformatics/btu153. Epub 2014 Mar 18. PMID: 24642063. +- [Krona](https://doi.org/10.1186/1471-2105-12-385) -* [SAMtools](https://doi.org/10.1093/bioinformatics/btp352) - > Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., … 1000 Genome Project Data Processing Subgroup. (2009). The Sequence Alignment/Map format and SAMtools. Bioinformatics , 25(16), 2078–2079. doi: 10.1093/bioinformatics/btp352. + > Ondov, B. D., Bergman, N. H., & Phillippy, A. M. (2011). Interactive metagenomic visualization in a Web browser. BMC bioinformatics, 12(1), 1-10. doi: 10.1186/1471-2105-12-385. -* [SPAdes](https://doi.org/10.1101/gr.213959.116) - > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. +- [MaxBin2](https://doi.org/10.1093/bioinformatics/btv638) -* [PyDamage](https://doi.org/10.7717/peerj.11845) - > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 + > Yu-Wei, W., Simmons, B. A. & Singer, S. W. (2015) MaxBin 2.0: An Automated Binning Algorithm to Recover Genomes from Multiple Metagenomic Datasets. Bioinformatics 32 (4): 605–7. doi: 10.1093/bioinformatics/btv638. + +- [MEGAHIT](https://doi.org/10.1016/j.ymeth.2016.02.020) + + > Li, D., Luo, R., Liu, C. M., Leung, C. M., Ting, H. F., Sadakane, K., ... & Lam, T. W. (2016). MEGAHIT v1. 0: a fast and scalable metagenome assembler driven by advanced methodologies and community practices. Methods, 102, 3-11. doi: 10.1016/j.ymeth.2016.02.020. + +- [MetaBAT2](https://doi.org/10.7717/peerj.7359) + + > Kang, D. D., Li, F., Kirton, E., Thomas, A., Egan, R., An, H., & Wang, Z. (2019). MetaBAT 2: an adaptive binning algorithm for robust and efficient genome reconstruction from metagenome assemblies. PeerJ, 7, e7359. doi: 10.7717/peerj.7359. + +- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) + + > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. + +- [NanoLyse](https://doi.org/10.1093/bioinformatics/bty149) + + > De Coster, W., D’Hert, S., Schultz, D. T., Cruts, M., & Van Broeckhoven, C. (2018). NanoPack: visualizing and processing long-read sequencing data. Bioinformatics, 34(15), 2666-2669. doi: 10.1093/bioinformatics/bty149. + +- [NanoPlot](https://doi.org/10.1093/bioinformatics/bty149) + + > De Coster, W., D’Hert, S., Schultz, D. T., Cruts, M., & Van Broeckhoven, C. (2018). NanoPack: visualizing and processing long-read sequencing data. Bioinformatics, 34(15), 2666-2669. doi: 10.1093/bioinformatics/bty149. + +- [Porechop](https://github.com/rrwick/Porechop) + +- [Prodigal](https://pubmed.ncbi.nlm.nih.gov/20211023/) + + > Hyatt D, Chen GL, Locascio PF, Land ML, Larimer FW, Hauser LJ. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinformatics. 2010 Mar 8;11:119. doi: 10.1186/1471-2105-11-119. PMID: 20211023; PMCID: PMC2848648. + +- [Prokka](https://pubmed.ncbi.nlm.nih.gov/24642063/) + + > Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014 Jul 15;30(14):2068-9. doi: 10.1093/bioinformatics/btu153. Epub 2014 Mar 18. PMID: 24642063. + +- [SAMtools](https://doi.org/10.1093/bioinformatics/btp352) + + > Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., … 1000 Genome Project Data Processing Subgroup. (2009). The Sequence Alignment/Map format and SAMtools. Bioinformatics , 25(16), 2078–2079. doi: 10.1093/bioinformatics/btp352. + +- [SPAdes](https://doi.org/10.1101/gr.213959.116) + + > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. + +- [PyDamage](https://doi.org/10.7717/peerj.11845) + > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 ## Data -* [Full-size test data](https://doi.org/10.1038/s41587-019-0191-2) - > Bertrand, D., Shaw, J., Kalathiyappan, M., Ng, A. H. Q., Kumar, M. S., Li, C., ... & Nagarajan, N. (2019). Hybrid metagenomic assembly enables high-resolution analysis of resistance determinants and mobile elements in human microbiomes. Nature biotechnology, 37(8), 937-944. doi: 10.1038/s41587-019-0191-2. +- [Full-size test data](https://doi.org/10.1038/s41587-019-0191-2) + > Bertrand, D., Shaw, J., Kalathiyappan, M., Ng, A. H. Q., Kumar, M. S., Li, C., ... & Nagarajan, N. (2019). Hybrid metagenomic assembly enables high-resolution analysis of resistance determinants and mobile elements in human microbiomes. Nature biotechnology, 37(8), 937-944. doi: 10.1038/s41587-019-0191-2. ## Software packaging/containerisation tools diff --git a/README.md b/README.md index 9178cbd8..df510fac 100644 --- a/README.md +++ b/README.md @@ -32,12 +32,12 @@ On release, automated continuous integration tests run the pipeline on a full-si By default, the pipeline currently performs the following: it supports both short and long reads, quality trims the reads and adapters with [fastp](https://github.com/OpenGene/fastp) and [Porechop](https://github.com/rrwick/Porechop), and performs basic QC with [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/). The pipeline then: -* assigns taxonomy to reads using [Centrifuge](https://ccb.jhu.edu/software/centrifuge/) and/or [Kraken2](https://github.com/DerrickWood/kraken2/wiki) -* performs assembly using [MEGAHIT](https://github.com/voutcn/megahit) and [SPAdes](http://cab.spbu.ru/software/spades/), and checks their quality using [Quast](http://quast.sourceforge.net/quast) -* (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) -* predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) -* performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/) and/or with [MaxBin2](https://sourceforge.net/projects/maxbin2/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) -* assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) +- assigns taxonomy to reads using [Centrifuge](https://ccb.jhu.edu/software/centrifuge/) and/or [Kraken2](https://github.com/DerrickWood/kraken2/wiki) +- performs assembly using [MEGAHIT](https://github.com/voutcn/megahit) and [SPAdes](http://cab.spbu.ru/software/spades/), and checks their quality using [Quast](http://quast.sourceforge.net/quast) +- (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) +- predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) +- performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/) and/or with [MaxBin2](https://sourceforge.net/projects/maxbin2/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) +- assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. @@ -62,15 +62,15 @@ Furthermore, the pipeline creates various reports in the results directory speci 4. Start running your own analysis! - ```console - nextflow run nf-core/mag -profile --input '*_R{1,2}.fastq.gz' --outdir - ``` + ```console + nextflow run nf-core/mag -profile --input '*_R{1,2}.fastq.gz' --outdir + ``` - or + or - ```console - nextflow run nf-core/mag -profile --input samplesheet.csv --outdir - ``` + ```console + nextflow run nf-core/mag -profile --input samplesheet.csv --outdir + ``` See [usage docs](https://nf-co.re/mag/usage) and [parameter docs](https://nf-co.re/mag/parameters) for all of the available options when running the pipeline. @@ -92,15 +92,15 @@ Long read processing was inspired by [caspargross/HybridAssembly](https://github We thank the following people for their extensive assistance in the development of this pipeline: -* [Alexander Peltzer](https://github.com/apeltzer) -* [Antonia Schuster](https://github.com/antoniaschuster) -* [Phil Ewels](https://github.com/ewels) -* [Gisela Gabernet](https://github.com/ggabernet) -* [Harshil Patel](https://github.com/drpatelh) -* [Johannes Alneberg](https://github.com/alneberg) -* [Maxime Borry](https://github.com/maxibor) -* [Maxime Garcia](https://github.com/MaxUlysse) -* [Michael L Heuer](https://github.com/heuermh) +- [Alexander Peltzer](https://github.com/apeltzer) +- [Antonia Schuster](https://github.com/antoniaschuster) +- [Phil Ewels](https://github.com/ewels) +- [Gisela Gabernet](https://github.com/ggabernet) +- [Harshil Patel](https://github.com/drpatelh) +- [Johannes Alneberg](https://github.com/alneberg) +- [Maxime Borry](https://github.com/maxibor) +- [Maxime Garcia](https://github.com/MaxUlysse) +- [Michael L Heuer](https://github.com/heuermh) ## Contributions and Support diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index fa57bcb4..f146830d 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,47 +1,46 @@ report_comment: > - This report has been generated by the nf-core/mag - analysis pipeline. For information about how to interpret these results, please see the - documentation. + This report has been generated by the nf-core/mag + analysis pipeline. For information about how to interpret these results, please see the + documentation. report_section_order: - software_versions: - order: -1000 - nf-core-mag-summary: - order: -1001 + software_versions: + order: -1000 + nf-core-mag-summary: + order: -1001 export_plots: true -data_format: 'yaml' +data_format: "yaml" top_modules: - - 'fastqc': - name: 'FastQC: raw reads' - path_filters_exclude: - - '*trimmed*' - - custom_content - - 'fastqc': - name: 'FastQC: after preprocessing' - info: 'After trimming and, if requested, contamination removal.' - path_filters: - - '*trimmed*' - - 'busco': - info: 'assesses genome assembly and annotation completeness with Benchmarking Universal Single-Copy Orthologs. In case BUSCO''s automated lineage selection was used, only generic results for the selected domain are shown and only for genome bins and kept, unbinned contigs for which the BUSCO analysis was successfull, i.e. not for contigs for which no BUSCO genes could be found. Bins for which a specific virus lineage was selected are also not shown.' - - 'quast' - + - "fastqc": + name: "FastQC: raw reads" + path_filters_exclude: + - "*trimmed*" + - custom_content + - "fastqc": + name: "FastQC: after preprocessing" + info: "After trimming and, if requested, contamination removal." + path_filters: + - "*trimmed*" + - "busco": + info: "assesses genome assembly and annotation completeness with Benchmarking Universal Single-Copy Orthologs. In case BUSCO's automated lineage selection was used, only generic results for the selected domain are shown and only for genome bins and kept, unbinned contigs for which the BUSCO analysis was successfull, i.e. not for contigs for which no BUSCO genes could be found. Bins for which a specific virus lineage was selected are also not shown." + - "quast" custom_data: - host_removal: - file_format: 'tsv' - section_name: 'Bowtie 2: host read removal' - description: 'Reads are mapped against the host reference sequence. Only reads that do not align (concordantly) are kept for further analysis.' - plot_type: 'bargraph' - pconfig: - id: 'host_removal_bargraph' - title: 'Bowtie 2: reads mapped against host reference' - ylab: '# Reads' + host_removal: + file_format: "tsv" + section_name: "Bowtie 2: host read removal" + description: "Reads are mapped against the host reference sequence. Only reads that do not align (concordantly) are kept for further analysis." + plot_type: "bargraph" + pconfig: + id: "host_removal_bargraph" + title: "Bowtie 2: reads mapped against host reference" + ylab: "# Reads" sp: - host_removal: - fn: 'host_removal_metrics.tsv' + host_removal: + fn: "host_removal_metrics.tsv" extra_fn_clean_exts: - - '.bowtie2' + - ".bowtie2" diff --git a/docs/output.md b/docs/output.md index 61488ec1..d8b13601 100644 --- a/docs/output.md +++ b/docs/output.md @@ -10,16 +10,16 @@ The directories listed below will be created in the results directory after the The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: -* [Quality control](#quality-control) of input reads - trimming and contaminant removal -* [Taxonomic classification of trimmed reads](#taxonomic-classification-of-trimmed-reads) -* [Assembly](#assembly) of trimmed reads -* [Protein-coding gene prediction](#gene-prediction) of assemblies -* [Binning](#binning) of assembled contigs -* [Taxonomic classification of binned genomes](#taxonomic-classification-of-binned-genomes) -* [Genome annotation of binned genomes](#genome-annotation-of-binned-genomes) -* [Additional summary for binned genomes](#additional-summary-for-binned-genomes) -* [MultiQC](#multiqc) - aggregate report, describing results of the whole pipeline -* [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution +- [Quality control](#quality-control) of input reads - trimming and contaminant removal +- [Taxonomic classification of trimmed reads](#taxonomic-classification-of-trimmed-reads) +- [Assembly](#assembly) of trimmed reads +- [Protein-coding gene prediction](#gene-prediction) of assemblies +- [Binning](#binning) of assembled contigs +- [Taxonomic classification of binned genomes](#taxonomic-classification-of-binned-genomes) +- [Genome annotation of binned genomes](#genome-annotation-of-binned-genomes) +- [Additional summary for binned genomes](#additional-summary-for-binned-genomes) +- [MultiQC](#multiqc) - aggregate report, describing results of the whole pipeline +- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution Note that when specifying the parameter `--coassemble_group`, for the corresponding output filenames/directories of the assembly or downsteam processes the group ID, or more precisely the term `group-[group_id]`, will be used instead of the sample ID. @@ -36,9 +36,9 @@ FastQC is run for visualising the general quality metrics of the sequencing runs
Output files -* `QC_shortreads/fastqc/` - * `[sample]_[1/2]_fastqc.html`: FastQC report, containing quality metrics for your untrimmed raw fastq files - * `[sample].trimmed_[1/2]_fastqc.html`: FastQC report, containing quality metrics for trimmed and, if specified, filtered read files +- `QC_shortreads/fastqc/` + - `[sample]_[1/2]_fastqc.html`: FastQC report, containing quality metrics for your untrimmed raw fastq files + - `[sample].trimmed_[1/2]_fastqc.html`: FastQC report, containing quality metrics for trimmed and, if specified, filtered read files
@@ -51,9 +51,9 @@ FastQC is run for visualising the general quality metrics of the sequencing runs
Output files -* `QC_shortreads/fastp/[sample]/` - * `fastp.html`: Interactive report - * `fastp.json`: Report in json format +- `QC_shortreads/fastp/[sample]/` + - `fastp.html`: Interactive report + - `fastp.json`: Report in json format
@@ -64,8 +64,8 @@ The pipeline uses bowtie2 to map the reads against PhiX and removes mapped reads
Output files -* `QC_shortreads/remove_phix/` - * `[sample].phix_removed.bowtie2.log`: Contains a brief log file indicating how many reads have been retained. +- `QC_shortreads/remove_phix/` + - `[sample].phix_removed.bowtie2.log`: Contains a brief log file indicating how many reads have been retained.
@@ -76,8 +76,8 @@ The pipeline uses bowtie2 to map short reads against the host reference genome s
Output files -* `QC_shortreads/remove_host/` - * `[sample].host_removed.bowtie2.log`: Contains the bowtie2 log file indicating how many reads have been mapped as well as a file listing the read ids of discarded reads. +- `QC_shortreads/remove_host/` + - `[sample].host_removed.bowtie2.log`: Contains the bowtie2 log file indicating how many reads have been mapped as well as a file listing the read ids of discarded reads.
@@ -88,8 +88,8 @@ The pipeline uses Nanolyse to map the reads against the Lambda phage and removes
Output files -* `QC_longreads/NanoLyse/` - * `[sample]_nanolyse.log`: Contains a brief log file indicating how many reads have been retained. +- `QC_longreads/NanoLyse/` + - `[sample]_nanolyse.log`: Contains a brief log file indicating how many reads have been retained.
@@ -109,9 +109,9 @@ NanoPlot is used to calculate various metrics and plots about the quality and le
Output files -* `QC_longreads/NanoPlot/[sample]/` - * `raw_*.[png/html/txt]`: Plots and reports for raw data - * `filtered_*.[png/html/txt]`: Plots and reports for filtered data +- `QC_longreads/NanoPlot/[sample]/` + - `raw_*.[png/html/txt]`: Plots and reports for raw data + - `filtered_*.[png/html/txt]`: Plots and reports for filtered data
@@ -124,9 +124,9 @@ Kraken2 classifies reads using a k-mer based approach as well as assigns taxonom
Output files -* `Taxonomy/kraken2/[sample]/` - * `kraken2.report`: Classification in the Kraken report format. See the [kraken2 manual](https://github.com/DerrickWood/kraken2/wiki/Manual#output-formats) for more details - * `taxonomy.krona.html`: Interactive pie chart produced by [KronaTools](https://github.com/marbl/Krona/wiki) +- `Taxonomy/kraken2/[sample]/` + - `kraken2.report`: Classification in the Kraken report format. See the [kraken2 manual](https://github.com/DerrickWood/kraken2/wiki/Manual#output-formats) for more details + - `taxonomy.krona.html`: Interactive pie chart produced by [KronaTools](https://github.com/marbl/Krona/wiki)
@@ -139,10 +139,10 @@ More information on the [Centrifuge](https://ccb.jhu.edu/software/centrifuge/) w
Output files -* `Taxonomy/centrifuge/[sample]/` - * `report.txt`: Tab-delimited result file. See the [centrifuge manual](https://ccb.jhu.edu/software/centrifuge/manual.shtml#centrifuge-classification-output) for information about the fields - * `kreport.txt`: Classification in the Kraken report format. See the [kraken2 manual](https://github.com/DerrickWood/kraken2/wiki/Manual#output-formats) for more details - * `taxonomy.krona.html`: Interactive pie chart produced by [KronaTools](https://github.com/marbl/Krona/wiki) +- `Taxonomy/centrifuge/[sample]/` + - `report.txt`: Tab-delimited result file. See the [centrifuge manual](https://ccb.jhu.edu/software/centrifuge/manual.shtml#centrifuge-classification-output) for information about the fields + - `kreport.txt`: Classification in the Kraken report format. See the [kraken2 manual](https://github.com/DerrickWood/kraken2/wiki/Manual#output-formats) for more details + - `taxonomy.krona.html`: Interactive pie chart produced by [KronaTools](https://github.com/marbl/Krona/wiki)
@@ -157,12 +157,12 @@ Trimmed (short) reads are assembled with both megahit and SPAdes. Hybrid assembl
Output files -* `Assembly/MEGAHIT/` - * `[sample/group].contigs.fa.gz`: Compressed metagenome assembly in fasta format - * `[sample/group].log`: Log file - * `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs - * `MEGAHIT-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. - * `MEGAHIT-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap"). +- `Assembly/MEGAHIT/` + - `[sample/group].contigs.fa.gz`: Compressed metagenome assembly in fasta format + - `[sample/group].log`: Log file + - `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs + - `MEGAHIT-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. + - `MEGAHIT-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap").
@@ -173,14 +173,14 @@ Trimmed (short) reads are assembled with both megahit and SPAdes. Hybrid assembl
Output files -* `Assembly/SPAdes/` - * `[sample/group]_scaffolds.fasta.gz`: Compressed assembled scaffolds in fasta format - * `[sample/group]_graph.gfa.gz`: Compressed assembly graph in gfa format - * `[sample/group]_contigs.fasta.gz`: Compressed assembled contigs in fasta format - * `[sample/group].log`: Log file - * `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs - * `SPAdes-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. - * `SPAdes-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap"). +- `Assembly/SPAdes/` + - `[sample/group]_scaffolds.fasta.gz`: Compressed assembled scaffolds in fasta format + - `[sample/group]_graph.gfa.gz`: Compressed assembly graph in gfa format + - `[sample/group]_contigs.fasta.gz`: Compressed assembled contigs in fasta format + - `[sample/group].log`: Log file + - `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs + - `SPAdes-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. + - `SPAdes-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap").
@@ -191,14 +191,14 @@ SPAdesHybrid is a part of the [SPAdes](http://cab.spbu.ru/software/spades/) soft
Output files -* `Assembly/SPAdesHybrid/` - * `[sample/group]_scaffolds.fasta.gz`: Compressed assembled scaffolds in fasta format - * `[sample/group]_graph.gfa.gz`: Compressed assembly graph in gfa format - * `[sample/group]_contigs.fasta.gz`: Compressed assembled contigs in fasta format - * `[sample/group].log`: Log file - * `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs - * `SPAdesHybrid-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. - * `SPAdesHybrid-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap"). +- `Assembly/SPAdesHybrid/` + - `[sample/group]_scaffolds.fasta.gz`: Compressed assembled scaffolds in fasta format + - `[sample/group]_graph.gfa.gz`: Compressed assembly graph in gfa format + - `[sample/group]_contigs.fasta.gz`: Compressed assembled contigs in fasta format + - `[sample/group].log`: Log file + - `QC/[sample/group]/`: Directory containing QUAST files and Bowtie2 mapping logs + - `SPAdesHybrid-[sample].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the sample that the metagenome was assembled from, only present if `--coassemble_group` is not set. + - `SPAdesHybrid-[sample/group]-[sampleToMap].bowtie2.log`: Bowtie2 log file indicating how many reads have been mapped from the respective sample ("sampleToMap").
@@ -209,19 +209,19 @@ SPAdesHybrid is a part of the [SPAdes](http://cab.spbu.ru/software/spades/) soft
Output files -* `Assembly/[assembler]/QC/[sample/group]/QUAST/` - * `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt - * `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) - * `quast.log`: QUAST log file - * `metaquast.log`: MetaQUAST log file - * `icarus.html`: Icarus main menu with links to interactive viewers - * `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest - * `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) - * `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs - * `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs - * `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. - * `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format - * `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor) +- `Assembly/[assembler]/QC/[sample/group]/QUAST/` + - `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt + - `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) + - `quast.log`: QUAST log file + - `metaquast.log`: MetaQUAST log file + - `icarus.html`: Icarus main menu with links to interactive viewers + - `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest + - `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) + - `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs + - `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs + - `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. + - `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format + - `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor)
@@ -232,11 +232,11 @@ Protein-coding genes are predicted for each assembly.
Output files -* `Prodigal/` - * `[sample/group].gff`: Gene Coordinates in GFF format - * `[sample/group].faa`: The protein translation file consists of all the proteins from all the sequences in multiple FASTA format. - * `[sample/group].fna`: Nucleotide sequences of the predicted proteins using the DNA alphabet, not mRNA (so you will see 'T' in the output and not 'U'). - * `[sample/group]_all.txt`: Information about start positions of genes. +- `Prodigal/` + - `[sample/group].gff`: Gene Coordinates in GFF format + - `[sample/group].faa`: The protein translation file consists of all the proteins from all the sequences in multiple FASTA format. + - `[sample/group].fna`: Nucleotide sequences of the predicted proteins using the DNA alphabet, not mRNA (so you will see 'T' in the output and not 'U'). + - `[sample/group]_all.txt`: Information about start positions of genes.
@@ -249,8 +249,8 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig
Output files -* `GenomeBinning/` - * `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads. +- `GenomeBinning/` + - `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads.
@@ -261,9 +261,9 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig
Output files -* `GenomeBinning/MetaBAT2/` - * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported +- `GenomeBinning/MetaBAT2/` + - `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + - `unbinned/[assembler]-[binner]-[sample/group].unbinned.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported
@@ -272,12 +272,12 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO.
Output files -* `GenomeBinning/MetaBAT2/discarded/` - * `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBAT2 - * `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBAT2 -* `GenomeBinning/MetaBAT2/unbinned/discarded/` - * `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. - * `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. +- `GenomeBinning/MetaBAT2/discarded/` + - `*.lowDepth.fa.gz`: Low depth contigs that are filtered by MetaBAT2 + - `*.tooShort.fa.gz`: Too short contigs that are filtered by MetaBAT2 +- `GenomeBinning/MetaBAT2/unbinned/discarded/` + - `*.unbinned.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. + - `*.unbinned.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file.
@@ -292,9 +292,9 @@ Files in these two folders contain all contigs of an assembly.
Output files -* `GenomeBinning/MaxBin2/` - * `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - * `unbinned/[assembler]-[binner]-[sample/group].noclass.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. +- `GenomeBinning/MaxBin2/` + - `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + - `unbinned/[assembler]-[binner]-[sample/group].noclass.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported.
@@ -303,11 +303,11 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO.
Output files -* `GenomeBinning/MaxBin2/discarded/` - * `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 -* `GenomeBinning/MaxBin2/unbinned/discarded/` - * `*.noclass.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. - * `*.noclass.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. +- `GenomeBinning/MaxBin2/discarded/` + - `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 +- `GenomeBinning/MaxBin2/unbinned/discarded/` + - `*.noclass.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. + - `*.noclass.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file.
@@ -322,9 +322,9 @@ For each genome bin the median sequencing depth is computed based on the corresp
Output files -* `GenomeBinning/` - * `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - * `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. +- `GenomeBinning/` + - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. + - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`.
@@ -335,21 +335,21 @@ For each genome bin the median sequencing depth is computed based on the corresp
Output files -* `GenomeBinning/QC/QUAST/[assembler]-[bin]/` - * `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt - * `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) - * `quast.log`: QUAST log file - * `metaquast.log`: MetaQUAST log file - * `icarus.html`: Icarus main menu with links to interactive viewers - * `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest - * `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) - * `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs - * `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs - * `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. - * `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format - * `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor) -* `GenomeBinning/QC/` - * `quast_summary.tsv`: QUAST output for all bins summarized +- `GenomeBinning/QC/QUAST/[assembler]-[bin]/` + - `report.*`: QUAST report in various formats, such as html, pdf, tex, tsv, or txt + - `transposed_report.*`: QUAST report that has been transposed into wide format (tex, tsv, or txt) + - `quast.log`: QUAST log file + - `metaquast.log`: MetaQUAST log file + - `icarus.html`: Icarus main menu with links to interactive viewers + - `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest + - `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) + - `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs + - `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs + - `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. + - `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format + - `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor) +- `GenomeBinning/QC/` + - `quast_summary.tsv`: QUAST output for all bins summarized
@@ -360,14 +360,14 @@ For each genome bin the median sequencing depth is computed based on the corresp
Output files -* `GenomeBinning/QC/BUSCO/` - * `[assembler]-[bin]_busco.log`: Log file containing the standard output of BUSCO. - * `[assembler]-[bin]_busco.err`: File containing potential error messages returned from BUSCO. - * `short_summary.domain.[lineage].[assembler]-[bin].txt`: BUSCO summary of the results for the selected domain when run in automated lineage selection mode. Not available for bins for which a viral lineage was selected. - * `short_summary.specific_lineage.[lineage].[assembler]-[bin].txt`: BUSCO summary of the results in case a more specific lineage than the domain could be selected or for the lineage provided via `--busco_reference`. - * `[assembler]-[bin]_buscos.[lineage].fna.gz`: Nucleotide sequence of all identified BUSCOs for used lineages (domain or specific). - * `[assembler]-[bin]_buscos.[lineage].faa.gz`: Aminoacid sequence of all identified BUSCOs for used lineages (domain or specific). - * `[assembler]-[bin]_prodigal.gff`: Genes predicted with Prodigal. +- `GenomeBinning/QC/BUSCO/` + - `[assembler]-[bin]_busco.log`: Log file containing the standard output of BUSCO. + - `[assembler]-[bin]_busco.err`: File containing potential error messages returned from BUSCO. + - `short_summary.domain.[lineage].[assembler]-[bin].txt`: BUSCO summary of the results for the selected domain when run in automated lineage selection mode. Not available for bins for which a viral lineage was selected. + - `short_summary.specific_lineage.[lineage].[assembler]-[bin].txt`: BUSCO summary of the results in case a more specific lineage than the domain could be selected or for the lineage provided via `--busco_reference`. + - `[assembler]-[bin]_buscos.[lineage].fna.gz`: Nucleotide sequence of all identified BUSCOs for used lineages (domain or specific). + - `[assembler]-[bin]_buscos.[lineage].faa.gz`: Aminoacid sequence of all identified BUSCOs for used lineages (domain or specific). + - `[assembler]-[bin]_prodigal.gff`: Genes predicted with Prodigal.
@@ -376,9 +376,9 @@ If the parameter `--save_busco_reference` is set, additionally the used BUSCO li
Output files -* `GenomeBinning/QC/BUSCO/` - * `busco_downloads/`: All files and lineage datasets downloaded by BUSCO when run in automated lineage selection mode. (Can currently not be used to reproduce analysis, see the [nf-core/mag website documentation](https://nf-co.re/mag/usage#reproducibility) how to achieve reproducible BUSCO results). - * `reference/*.tar.gz`: BUSCO reference lineage dataset that was provided via `--busco_reference`. +- `GenomeBinning/QC/BUSCO/` + - `busco_downloads/`: All files and lineage datasets downloaded by BUSCO when run in automated lineage selection mode. (Can currently not be used to reproduce analysis, see the [nf-core/mag website documentation](https://nf-co.re/mag/usage#reproducibility) how to achieve reproducible BUSCO results). + - `reference/*.tar.gz`: BUSCO reference lineage dataset that was provided via `--busco_reference`.
@@ -387,8 +387,8 @@ Besides the reference files or output files created by BUSCO, the following summ
Output files -* `GenomeBinning/QC/` - * `busco_summary.tsv`: A summary table of the BUSCO results, with % of marker genes found. If run in automated lineage selection mode, both the results for the selected domain and for the selected more specific lineage will be given, if available. +- `GenomeBinning/QC/` + - `busco_summary.tsv`: A summary table of the BUSCO results, with % of marker genes found. If run in automated lineage selection mode, both the results for the selected domain and for the selected more specific lineage will be given, if available.
@@ -401,15 +401,15 @@ Besides the reference files or output files created by BUSCO, the following summ
Output files -* `Taxonomy/CAT/[assembler]/` - * `[assembler]-[sample/group].ORF2LCA.names.txt.gz`: Tab-delimited files containing the lineage of each contig, with full lineage names - * `[assembler]-[sample/group].bin2classification.names.txt.gz`: Taxonomy classification of the genome bins, with full lineage names -* `Taxonomy/CAT/[assembler]/raw/` - * `[assembler]-[sample/group].concatenated.predicted_proteins.faa.gz`: Predicted protein sequences for each genome bin, in fasta format - * `[assembler]-[sample/group].concatenated.predicted_proteins.gff.gz`: Predicted protein features for each genome bin, in gff format - * `[assembler]-[sample/group].ORF2LCA.txt.gz`: Tab-delimited files containing the lineage of each contig - * `[assembler]-[sample/group].bin2classification.txt.gz`: Taxonomy classification of the genome bins - * `[assembler]-[sample/group].log`: Log files +- `Taxonomy/CAT/[assembler]/` + - `[assembler]-[sample/group].ORF2LCA.names.txt.gz`: Tab-delimited files containing the lineage of each contig, with full lineage names + - `[assembler]-[sample/group].bin2classification.names.txt.gz`: Taxonomy classification of the genome bins, with full lineage names +- `Taxonomy/CAT/[assembler]/raw/` + - `[assembler]-[sample/group].concatenated.predicted_proteins.faa.gz`: Predicted protein sequences for each genome bin, in fasta format + - `[assembler]-[sample/group].concatenated.predicted_proteins.gff.gz`: Predicted protein features for each genome bin, in gff format + - `[assembler]-[sample/group].ORF2LCA.txt.gz`: Tab-delimited files containing the lineage of each contig + - `[assembler]-[sample/group].bin2classification.txt.gz`: Taxonomy classification of the genome bins + - `[assembler]-[sample/group].log`: Log files
@@ -418,7 +418,7 @@ If the parameters `--cat_db_generate` and `--save_cat_db` are set, additionally
Output files -* `Taxonomy/CAT/CAT_prepare_*.tar.gz`: Generated and used CAT database. +- `Taxonomy/CAT/CAT_prepare_*.tar.gz`: Generated and used CAT database.
@@ -429,15 +429,15 @@ If the parameters `--cat_db_generate` and `--save_cat_db` are set, additionally
Output files -* `Taxonomy/GTDB-Tk/[assembler]/[sample/group]/` - * `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.summary.tsv`: Classifications for bacterial and archaeal genomes (see the [GTDB-Tk documentation for details](https://ecogenomics.github.io/GTDBTk/files/summary.tsv.html). - * `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.classify.tree.gz`: Reference tree in Newick format containing query genomes placed with pplacer. - * `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.markers_summary.tsv`: A summary of unique, duplicated, and missing markers within the 120 bacterial marker set, or the 122 archaeal marker set for each submitted genome. - * `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.msa.fasta.gz`: FASTA file containing MSA of submitted and reference genomes. - * `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.filtered.tsv`: A list of genomes with an insufficient number of amino acids in MSA. - * `gtdbtk.[assembler]-[sample/group].*.log`: Log files. - * `gtdbtk.[assembler]-[sample/group].failed_genomes.tsv`: A list of genomes for which the GTDB-Tk analysis failed, e.g. because Prodigal could not detect any genes. -* `Taxonomy/GTDB-Tk/gtdbtk_summary.tsv`: A summary table of the GTDB-Tk classification results for all bins, also containing bins which were discarded based on the BUSCO QC, which were filtered out by GTDB-Tk ((listed in `*.filtered.tsv`) or for which the analysis failed (listed in `*.failed_genomes.tsv`). +- `Taxonomy/GTDB-Tk/[assembler]/[sample/group]/` + - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.summary.tsv`: Classifications for bacterial and archaeal genomes (see the [GTDB-Tk documentation for details](https://ecogenomics.github.io/GTDBTk/files/summary.tsv.html). + - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.classify.tree.gz`: Reference tree in Newick format containing query genomes placed with pplacer. + - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.markers_summary.tsv`: A summary of unique, duplicated, and missing markers within the 120 bacterial marker set, or the 122 archaeal marker set for each submitted genome. + - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.msa.fasta.gz`: FASTA file containing MSA of submitted and reference genomes. + - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.filtered.tsv`: A list of genomes with an insufficient number of amino acids in MSA. + - `gtdbtk.[assembler]-[sample/group].*.log`: Log files. + - `gtdbtk.[assembler]-[sample/group].failed_genomes.tsv`: A list of genomes for which the GTDB-Tk analysis failed, e.g. because Prodigal could not detect any genes. +- `Taxonomy/GTDB-Tk/gtdbtk_summary.tsv`: A summary table of the GTDB-Tk classification results for all bins, also containing bins which were discarded based on the BUSCO QC, which were filtered out by GTDB-Tk ((listed in `*.filtered.tsv`) or for which the analysis failed (listed in `*.failed_genomes.tsv`).
@@ -450,19 +450,19 @@ Whole genome annotation is the process of identifying features of interest in a
Output files -* `Prokka/[assembler]/[bin]/` - * `[bin].gff`: annotation in GFF3 format, containing both sequences and annotations - * `[bin].gbk`: annotation in GenBank format, containing both sequences and annotations - * `[bin].fna`: nucleotide FASTA file of the input contig sequences - * `[bin].faa`: protein FASTA file of the translated CDS sequences - * `[bin].ffn`: nucleotide FASTA file of all the prediction transcripts (CDS, rRNA, tRNA, tmRNA, misc_RNA) - * `[bin].sqn`: an ASN1 format "Sequin" file for submission to Genbank - * `[bin].fsa`: nucleotide FASTA file of the input contig sequences, used by "tbl2asn" to create the .sqn file - * `[bin].tbl`: feature Table file, used by "tbl2asn" to create the .sqn file - * `[bin].err`: unacceptable annotations - the NCBI discrepancy report. - * `[bin].log`: contains all the output that Prokka produced during its run - * `[bin].txt`: statistics relating to the annotated features found - * `[bin].tsv`: tab-separated file of all features (locus_tag, ftype, len_bp, gene, EC_number, COG, product) +- `Prokka/[assembler]/[bin]/` + - `[bin].gff`: annotation in GFF3 format, containing both sequences and annotations + - `[bin].gbk`: annotation in GenBank format, containing both sequences and annotations + - `[bin].fna`: nucleotide FASTA file of the input contig sequences + - `[bin].faa`: protein FASTA file of the translated CDS sequences + - `[bin].ffn`: nucleotide FASTA file of all the prediction transcripts (CDS, rRNA, tRNA, tmRNA, misc_RNA) + - `[bin].sqn`: an ASN1 format "Sequin" file for submission to Genbank + - `[bin].fsa`: nucleotide FASTA file of the input contig sequences, used by "tbl2asn" to create the .sqn file + - `[bin].tbl`: feature Table file, used by "tbl2asn" to create the .sqn file + - `[bin].err`: unacceptable annotations - the NCBI discrepancy report. + - `[bin].log`: contains all the output that Prokka produced during its run + - `[bin].txt`: statistics relating to the annotated features found + - `[bin].tsv`: tab-separated file of all features (locus_tag, ftype, len_bp, gene, EC_number, COG, product)
@@ -471,7 +471,7 @@ Whole genome annotation is the process of identifying features of interest in a
Output files -* `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. +- `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated.
@@ -484,26 +484,26 @@ Whole genome annotation is the process of identifying features of interest in a
Output files -* `Ancient_DNA/pydamage/analyze` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) -* `Ancient_DNA/pydamage/filter` - * `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) +- `Ancient_DNA/pydamage/analyze` + - `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage raw result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html) +- `Ancient_DNA/pydamage/filter` + - `[sample/group]/pydamage_results/pydamage_results.csv`: PyDamage filtered result tabular file in `.csv` format. Format described here: [pydamage.readthedocs.io/en/0.62/output.html](https://pydamage.readthedocs.io/en/0.62/output.html)
### `variant_calling` -Because of aDNA damage, *de novo* assemblers sometimes struggle to call a correct consensus on the contig sequence. To avoid this situation, the consensus is re-called with a variant calling software using the reads aligned back to the contigs +Because of aDNA damage, _de novo_ assemblers sometimes struggle to call a correct consensus on the contig sequence. To avoid this situation, the consensus is re-called with a variant calling software using the reads aligned back to the contigs
Output files -* `variant_calling/consensus` - * `[sample/group].fa`: contigs sequence with re-called consensus from read-to-contig alignment -* `variant_calling/unfiltered` - * `[sample/group].vcf.gz`: raw variant calls of the reads aligned back to the contigs. -* `variant_calling/filtered` - * `[sample/group].filtered.vcf.gz`: quality filtered variant calls of the reads aligned back to the contigs. +- `variant_calling/consensus` + - `[sample/group].fa`: contigs sequence with re-called consensus from read-to-contig alignment +- `variant_calling/unfiltered` + - `[sample/group].vcf.gz`: raw variant calls of the reads aligned back to the contigs. +- `variant_calling/filtered` + - `[sample/group].filtered.vcf.gz`: quality filtered variant calls of the reads aligned back to the contigs.
@@ -528,9 +528,9 @@ Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQ
Output files -* `pipeline_info/` - * Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. - * Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. +- `pipeline_info/` + - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. + - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline.
diff --git a/docs/usage.md b/docs/usage.md index 96022028..ac4bab6e 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -20,13 +20,13 @@ This input method only works with short read data and will assign all files to t Please note the following additional requirements: -* Files names must be unique -* Valid file extensions: `.fastq.gz`, `.fq.gz` (files must be compressed) -* The path must be enclosed in quotes -* The path must have at least one `*` wildcard character -* When using the pipeline with paired end data, the path must use `{1,2}` notation to specify read pairs -* To run single-end data you must additionally specify `--single_end` -* If left unspecified, a default pattern is used: `data/*{1,2}.fastq.gz` +- Files names must be unique +- Valid file extensions: `.fastq.gz`, `.fq.gz` (files must be compressed) +- The path must be enclosed in quotes +- The path must have at least one `*` wildcard character +- When using the pipeline with paired end data, the path must use `{1,2}` notation to specify read pairs +- To run single-end data you must additionally specify `--single_end` +- If left unspecified, a default pattern is used: `data/*{1,2}.fastq.gz` ### Samplesheet input file @@ -55,14 +55,14 @@ sample2,0,data/sample2.fastq.gz,, Please note the following requirements: -* 5 comma-seperated columns -* Valid file extension: `.csv` -* Must contain the header `sample,group,short_reads_1,short_reads_2,long_reads` -* Sample IDs must be unique -* FastQ files must be compressed (`.fastq.gz`, `.fq.gz`) -* `long_reads` can only be provided in combination with paired-end short read data -* Within one samplesheet either only single-end or only paired-end reads can be specified -* If single-end reads are specified, the command line parameter `--single_end` must be specified as well +- 5 comma-seperated columns +- Valid file extension: `.csv` +- Must contain the header `sample,group,short_reads_1,short_reads_2,long_reads` +- Sample IDs must be unique +- FastQ files must be compressed (`.fastq.gz`, `.fq.gz`) +- `long_reads` can only be provided in combination with paired-end short read data +- Within one samplesheet either only single-end or only paired-end reads can be specified +- If single-end reads are specified, the command line parameter `--single_end` must be specified as well Again, by default, the group information is only used to compute co-abundances for the binning step, but not for group-wise co-assembly (see the parameter docs for [`--coassemble_group`](https://nf-co.re/mag/parameters#coassemble_group) and [`--binning_map_mode`](https://nf-co.re/mag/parameters#binning_map_mode) for more information about how this group information can be used). @@ -133,21 +133,21 @@ They are loaded in sequence, so later profiles can overwrite earlier profiles. If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended. -* `docker` - * A generic configuration profile to be used with [Docker](https://docker.com/) -* `singularity` - * A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/) -* `podman` - * A generic configuration profile to be used with [Podman](https://podman.io/) -* `shifter` - * A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) -* `charliecloud` - * A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) -* `conda` - * A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud. -* `test`, `test_hybrid`, `test_host_rm`, `test_hybrid_host_rm`, `test_busco_auto` - * Profiles with a complete configuration for automated testing - * Includes links to test data so needs no other parameters +- `docker` + - A generic configuration profile to be used with [Docker](https://docker.com/) +- `singularity` + - A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/) +- `podman` + - A generic configuration profile to be used with [Podman](https://podman.io/) +- `shifter` + - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) +- `charliecloud` + - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) +- `conda` + - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud. +- `test`, `test_hybrid`, `test_host_rm`, `test_hybrid_host_rm`, `test_busco_auto` + - Profiles with a complete configuration for automated testing + - Includes links to test data so needs no other parameters ### `-resume` @@ -286,4 +286,4 @@ NXF_OPTS='-Xms1g -Xmx4g' MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: -[Characteristic patterns of ancient DNA (aDNA) damage]((https://doi.org/10.1073/pnas.0704665104)), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. +[Characteristic patterns of ancient DNA (aDNA) damage](<(https://doi.org/10.1073/pnas.0704665104)>), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. diff --git a/modules.json b/modules.json index 4ed105fd..b11ea412 100644 --- a/modules.json +++ b/modules.json @@ -7,7 +7,7 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "bcftools/index": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "bcftools/view": { "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" @@ -16,13 +16,13 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "fastp": { - "git_sha": "e751e5040af57e1b4e06ed4e0f3efe6de25c1683" + "git_sha": "d0a1cbb703a130c19f6796c3fce24fbe7dfce789" }, "fastqc": { "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "freebayes": { - "git_sha": "7026122ec8696016b0b81496d9d39a7918cdb5b2" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "gunzip": { "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" @@ -31,26 +31,26 @@ "git_sha": "b78a4a456762a4c59fd5023e70f36a27f76d4a97" }, "metabat2/jgisummarizebamcontigdepths": { - "git_sha": "9d0cad583b9a71a6509b754fdf589cbfbed08961" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "metabat2/metabat2": { - "git_sha": "be8528998134132c6f4823f627bbb0aec6a51747" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "prodigal": { - "git_sha": "a45e03016632fe7dc2f48bfa73d9642c7f03e035" + "git_sha": "fcb1dce7b6f1563bdcdb70fdbe235cc2f9fed62e" }, "prokka": { - "git_sha": "d473a247d2e0c619b0df877ea19d9a5a98c8e3c8" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "pydamage/analyze": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "pydamage/filter": { - "git_sha": "20d8250d9f39ddb05dfb437603aaf99b5c0b2b41" + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "samtools/faidx": { "git_sha": "1ad73f1b2abdea9398680d6d20014838135c9a35" } } } -} +} \ No newline at end of file diff --git a/modules/nf-core/modules/bcftools/index/main.nf b/modules/nf-core/modules/bcftools/index/main.nf index 8f40c683..548a9277 100644 --- a/modules/nf-core/modules/bcftools/index/main.nf +++ b/modules/nf-core/modules/bcftools/index/main.nf @@ -2,10 +2,10 @@ process BCFTOOLS_INDEX { tag "$meta.id" label 'process_low' - conda (params.enable_conda ? 'bioconda::bcftools=1.13' : null) + conda (params.enable_conda ? 'bioconda::bcftools=1.14' : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/bcftools:1.13--h3a49de5_0' : - 'quay.io/biocontainers/bcftools:1.13--h3a49de5_0' }" + 'https://depot.galaxyproject.org/singularity/bcftools:1.14--h88f3f91_0' : + 'quay.io/biocontainers/bcftools:1.14--h88f3f91_0' }" input: tuple val(meta), path(vcf) @@ -13,11 +13,14 @@ process BCFTOOLS_INDEX { output: tuple val(meta), path("*.csi"), optional:true, emit: csi tuple val(meta), path("*.tbi"), optional:true, emit: tbi - path "versions.yml" , emit: version + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" """ bcftools \\ diff --git a/modules/nf-core/modules/bcftools/index/meta.yml b/modules/nf-core/modules/bcftools/index/meta.yml index 0d5dd3ef..b883fa5f 100644 --- a/modules/nf-core/modules/bcftools/index/meta.yml +++ b/modules/nf-core/modules/bcftools/index/meta.yml @@ -13,7 +13,7 @@ tools: documentation: https://samtools.github.io/bcftools/howtos/index.html tool_dev_url: https://github.com/samtools/bcftools doi: "10.1093/gigascience/giab008" - licence: ['MIT', 'GPL-3.0-or-later'] + licence: ["MIT", "GPL-3.0-or-later"] input: - meta: diff --git a/modules/nf-core/modules/fastp/main.nf b/modules/nf-core/modules/fastp/main.nf index a406036a..5c9e3b83 100644 --- a/modules/nf-core/modules/fastp/main.nf +++ b/modules/nf-core/modules/fastp/main.nf @@ -13,7 +13,7 @@ process FASTP { val save_merged output: - tuple val(meta), path('*.trim.fastq.gz') , emit: reads + tuple val(meta), path('*.trim.fastq.gz') , optional:true, emit: reads tuple val(meta), path('*.json') , emit: json tuple val(meta), path('*.html') , emit: html tuple val(meta), path('*.log') , emit: log @@ -21,6 +21,9 @@ process FASTP { tuple val(meta), path('*.fail.fastq.gz') , optional:true, emit: reads_fail tuple val(meta), path('*.merged.fastq.gz'), optional:true, emit: reads_merged + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' // Added soft-links to original fastqs for consistent naming in MultiQC diff --git a/modules/nf-core/modules/fastp/meta.yml b/modules/nf-core/modules/fastp/meta.yml index a1875faf..3274e41b 100644 --- a/modules/nf-core/modules/fastp/meta.yml +++ b/modules/nf-core/modules/fastp/meta.yml @@ -10,7 +10,7 @@ tools: A tool designed to provide fast all-in-one preprocessing for FastQ files. This tool is developed in C++ with multithreading supported to afford high performance. documentation: https://github.com/OpenGene/fastp doi: https://doi.org/10.1093/bioinformatics/bty560 - licence: ['MIT'] + licence: ["MIT"] input: - meta: type: map @@ -22,6 +22,12 @@ input: description: | List of input FastQ files of size 1 and 2 for single-end and paired-end data, respectively. + - save_trimmed_fail: + type: boolean + description: Specify true to save files that failed to pass trimming thresholds ending in `*.fail.fastq.gz` + - save_merged: + type: boolean + description: Specify true to save all merged reads to the a file ending in `*.merged.fastq.gz` output: - meta: diff --git a/modules/nf-core/modules/freebayes/main.nf b/modules/nf-core/modules/freebayes/main.nf index d462088a..73b1da96 100644 --- a/modules/nf-core/modules/freebayes/main.nf +++ b/modules/nf-core/modules/freebayes/main.nf @@ -8,10 +8,9 @@ process FREEBAYES { 'quay.io/biocontainers/freebayes:1.3.5--py38ha193a2f_3' }" input: - tuple val(meta), path(input_1), path(input_1_index), path(input_2), path(input_2_index) + tuple val(meta), path(input_1), path(input_1_index), path(input_2), path(input_2_index), path(target_bed) path fasta path fasta_fai - path targets path samples path populations path cnv @@ -20,11 +19,14 @@ process FREEBAYES { tuple val(meta), path("*.vcf.gz"), emit: vcf path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" def input = input_2 ? "${input_1} ${input_2}" : "${input_1}" - def targets_file = targets ? "--target ${targets}" : "" + def targets_file = target_bed ? "--target ${target_bed}" : "" def samples_file = samples ? "--samples ${samples}" : "" def populations_file = populations ? "--populations ${populations}" : "" def cnv_file = cnv ? "--cnv-map ${cnv}" : "" diff --git a/modules/nf-core/modules/freebayes/meta.yml b/modules/nf-core/modules/freebayes/meta.yml index abba1daa..cbbd297e 100644 --- a/modules/nf-core/modules/freebayes/meta.yml +++ b/modules/nf-core/modules/freebayes/meta.yml @@ -16,7 +16,7 @@ tools: documentation: https://github.com/freebayes/freebayes tool_dev_url: https://github.com/freebayes/freebayes doi: "arXiv:1207.3907" - licence: ['MIT'] + licence: ["MIT"] input: - meta: @@ -31,7 +31,11 @@ input: - input_index: type: file description: BAM/CRAM/SAM index file - pattern: "*.bam.bai" + pattern: "*.{bai,crai}" + - target_bed: + type: file + description: Optional - Limit analysis to targets listed in this BED-format FILE. + pattern: "*.bed" - fasta: type: file description: reference fasta file @@ -40,10 +44,6 @@ input: type: file description: reference fasta file index pattern: "*.{fa,fasta}.fai" - - targets: - type: file - description: Optional - Limit analysis to targets listed in this BED-format FILE. - pattern: "*.bed" - samples: type: file description: Optional - Limit analysis to samples listed (one per line) in the FILE. @@ -55,10 +55,10 @@ input: - cnv: type: file description: | - A copy number map BED file, which has either a sample-level ploidy: - sample_name copy_number - or a region-specific format: - seq_name start end sample_name copy_number + A copy number map BED file, which has either a sample-level ploidy: + sample_name copy_number + or a region-specific format: + seq_name start end sample_name copy_number pattern: "*.bed" output: diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf index 4a5869b6..7125eeb2 100644 --- a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/main.nf @@ -14,6 +14,9 @@ process METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { tuple val(meta), path("*.txt.gz"), emit: depth path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml index 351a4701..ff0ab40e 100644 --- a/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml +++ b/modules/nf-core/modules/metabat2/jgisummarizebamcontigdepths/meta.yml @@ -14,7 +14,7 @@ tools: documentation: https://bitbucket.org/berkeleylab/metabat/src/master/ tool_dev_url: https://bitbucket.org/berkeleylab/metabat/src/master/ doi: "10.7717/peerj.7359" - licence: ['BSD-3-clause-LBNL'] + licence: ["BSD-3-clause-LBNL"] input: - meta: diff --git a/modules/nf-core/modules/metabat2/metabat2/main.nf b/modules/nf-core/modules/metabat2/metabat2/main.nf index a8af0ae9..23ebe19a 100644 --- a/modules/nf-core/modules/metabat2/metabat2/main.nf +++ b/modules/nf-core/modules/metabat2/metabat2/main.nf @@ -18,6 +18,9 @@ process METABAT2_METABAT2 { tuple val(meta), path("bins/*.fa.gz") , optional:true , emit: fasta path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/nf-core/modules/metabat2/metabat2/meta.yml b/modules/nf-core/modules/metabat2/metabat2/meta.yml index 0ec07b02..04b8df4f 100644 --- a/modules/nf-core/modules/metabat2/metabat2/meta.yml +++ b/modules/nf-core/modules/metabat2/metabat2/meta.yml @@ -13,7 +13,7 @@ tools: documentation: https://bitbucket.org/berkeleylab/metabat/src/master/ tool_dev_url: https://bitbucket.org/berkeleylab/metabat/src/master/ doi: "10.7717/peerj.7359" - licence: ['BSD-3-clause-LBNL'] + licence: ["BSD-3-clause-LBNL"] input: - meta: @@ -28,8 +28,8 @@ input: - depth: type: file description: | - Optional text file listing the coverage per contig pre-generated - by metabat2_jgisummarizebamcontigdepths + Optional text file listing the coverage per contig pre-generated + by metabat2_jgisummarizebamcontigdepths pattern: "*.txt" output: @@ -63,7 +63,6 @@ output: description: cluster memberships as a matrix format. pattern: "*.tsv.gz" - authors: - "@maxibor" - "@jfy133" diff --git a/modules/nf-core/modules/prodigal/main.nf b/modules/nf-core/modules/prodigal/main.nf index 63248931..5768952b 100644 --- a/modules/nf-core/modules/prodigal/main.nf +++ b/modules/nf-core/modules/prodigal/main.nf @@ -18,6 +18,9 @@ process PRODIGAL { tuple val(meta), path("${prefix}_all.txt"), emit: all_gene_annotations path "versions.yml", emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/nf-core/modules/prodigal/meta.yml b/modules/nf-core/modules/prodigal/meta.yml index 3062ccce..8cb3d12e 100644 --- a/modules/nf-core/modules/prodigal/meta.yml +++ b/modules/nf-core/modules/prodigal/meta.yml @@ -5,9 +5,9 @@ keywords: tools: - prodigal: description: Prodigal (Prokaryotic Dynamic Programming Genefinding Algorithm) is a microbial (bacterial and archaeal) gene finding program - homepage: {https://github.com/hyattpd/Prodigal} - documentation: {https://github.com/hyattpd/prodigal/wiki} - tool_dev_url: {} + homepage: https://github.com/hyattpd/Prodigal + documentation: https://github.com/hyattpd/prodigal/wiki + tool_dev_url: https://github.com/hyattpd/Prodigal doi: "10.1186/1471-2105-11-119" licence: ["GPL v3"] diff --git a/modules/nf-core/modules/prokka/main.nf b/modules/nf-core/modules/prokka/main.nf index 551a17b9..3e46d1a1 100644 --- a/modules/nf-core/modules/prokka/main.nf +++ b/modules/nf-core/modules/prokka/main.nf @@ -27,6 +27,9 @@ process PROKKA { tuple val(meta), path("${prefix}/*.tsv"), emit: tsv path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/nf-core/modules/prokka/meta.yml b/modules/nf-core/modules/prokka/meta.yml index 87446694..7fc9e185 100644 --- a/modules/nf-core/modules/prokka/meta.yml +++ b/modules/nf-core/modules/prokka/meta.yml @@ -9,7 +9,7 @@ tools: description: Rapid annotation of prokaryotic genomes homepage: https://github.com/tseemann/prokka doi: "10.1093/bioinformatics/btu153" - licence: ['GPL v2'] + licence: ["GPL v2"] input: - meta: diff --git a/modules/nf-core/modules/pydamage/analyze/main.nf b/modules/nf-core/modules/pydamage/analyze/main.nf index c55616db..39e01fa1 100644 --- a/modules/nf-core/modules/pydamage/analyze/main.nf +++ b/modules/nf-core/modules/pydamage/analyze/main.nf @@ -14,9 +14,12 @@ process PYDAMAGE_ANALYZE { tuple val(meta), path("pydamage_results/pydamage_results.csv"), emit: csv path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" """ pydamage \\ analyze \\ diff --git a/modules/nf-core/modules/pydamage/analyze/meta.yml b/modules/nf-core/modules/pydamage/analyze/meta.yml index 918fbce9..09dd25eb 100644 --- a/modules/nf-core/modules/pydamage/analyze/meta.yml +++ b/modules/nf-core/modules/pydamage/analyze/meta.yml @@ -19,7 +19,7 @@ tools: homepage: https://github.com/maxibor/pydamage documentation: https://pydamage.readthedocs.io/ tool_dev_url: https://github.com/maxibor/pydamage - licence: ['GPL v3'] + licence: ["GPL v3"] input: - meta: diff --git a/modules/nf-core/modules/pydamage/filter/main.nf b/modules/nf-core/modules/pydamage/filter/main.nf index 2e0afac9..a748875a 100644 --- a/modules/nf-core/modules/pydamage/filter/main.nf +++ b/modules/nf-core/modules/pydamage/filter/main.nf @@ -14,9 +14,12 @@ process PYDAMAGE_FILTER { tuple val(meta), path("pydamage_results/pydamage_filtered_results.csv"), emit: csv path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" """ pydamage \\ diff --git a/modules/nf-core/modules/pydamage/filter/meta.yml b/modules/nf-core/modules/pydamage/filter/meta.yml index 706e38b0..c732ab9b 100644 --- a/modules/nf-core/modules/pydamage/filter/meta.yml +++ b/modules/nf-core/modules/pydamage/filter/meta.yml @@ -19,7 +19,7 @@ tools: homepage: https://github.com/maxibor/pydamage documentation: https://pydamage.readthedocs.io/ tool_dev_url: https://github.com/maxibor/pydamage - licence: ['GPL v3'] + licence: ["GPL v3"] input: - meta: From ab625b3cabe08fe1fc1128f6ec101054da6c550c Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:19:07 +0100 Subject: [PATCH 118/246] prettier again --- modules.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules.json b/modules.json index b11ea412..f3e7da69 100644 --- a/modules.json +++ b/modules.json @@ -53,4 +53,4 @@ } } } -} \ No newline at end of file +} From 8110363af934040726a350904dc83cfa45a02ade Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:21:12 +0100 Subject: [PATCH 119/246] fix freebayes input --- subworkflows/local/ancient_dna.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index e502c35a..4d2b0f62 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -21,6 +21,7 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { [], [], [], + [], [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) From 021e5dfc2212567bc96b58dddb497d6122d33403 Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:35:38 +0100 Subject: [PATCH 120/246] fix freebayes for real --- subworkflows/local/ancient_dna.nf | 4 +--- 1 file changed, 1 insertion(+), 3 deletions(-) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index 4d2b0f62..c59c02e8 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -15,13 +15,11 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { FAIDX(input.map { item -> [ item[0], item[1] ] }) input.join(FAIDX.out.fai) .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] - FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], []] }, + FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], [], []] }, freebayes_input.map { item -> item[1] }, freebayes_input.map { item -> item[4] }, [], [], - [], - [], [] ) BCFTOOLS_INDEX_PRE(FREEBAYES.out.vcf) From 5db23146e4cbe9ef354b6ce2af6749a502de2acb Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:36:02 +0100 Subject: [PATCH 121/246] fix README --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index df510fac..59fe2224 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,4 @@ -# ![nf-core/mag](docs/images/nf-core/mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core/mag_logo_dark.png#gh-dark-mode-only) +# ![nf-core/mag](docs/images/nf-core-mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core-mag_logo_dark.png#gh-dark-mode-only) [![GitHub Actions CI Status](https://github.com/nf-core/mag/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) From 02fbd2820c2983193559644f0127355950583aff Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 09:40:24 +0100 Subject: [PATCH 122/246] update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 14fa4c95..6ca8b907 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,7 +13,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Changed` -- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283) - Update to nf-core 2.3 `TEMPLATE` +- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283) - Update to nf-core 2.3.1 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint ### `Fixed` From ca0e4757eb3f11f19c6519a3f683154aafa9391c Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Thu, 24 Mar 2022 11:11:16 +0100 Subject: [PATCH 123/246] Apply suggestions from code review Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- CHANGELOG.md | 2 +- conf/modules.config | 2 +- docs/output.md | 2 -- workflows/mag.nf | 2 -- 4 files changed, 2 insertions(+), 6 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index d916cd96..2e8133f8 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,7 +10,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool -- [#264](https://github.com/nf-core/mag/pull/265) - Add AdapterRemoval2 as an alternative read trimmer +- [#284](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer ### `Changed` diff --git a/conf/modules.config b/conf/modules.config index 5566ed6b..4119fb6e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -35,7 +35,7 @@ process { "--cut_front", "--cut_tail", "--cut_mean_quality ${params.fastp_cut_mean_quality}", - "--length_required ${params.reads_minlength} " + "--length_required ${params.reads_minlength}" ].join(' ').trim() publishDir = [ path: { "${params.outdir}/QC_shortreads/fastp/${meta.id}" }, diff --git a/docs/output.md b/docs/output.md index e29274e4..3e4b4de7 100644 --- a/docs/output.md +++ b/docs/output.md @@ -59,8 +59,6 @@ FastQC is run for visualising the general quality metrics of the sequencing runs ### AdapterRemoval2 -[fastp](https://github.com/OpenGene/fastp) is a all-in-one fastq preprocessor for read/adapter trimming and quality control. It is used in this pipeline for trimming adapter sequences and discard low-quality reads. Its output is in the results folder and part of the MultiQC report. - [AdapterRemoval](https://adapterremoval.readthedocs.io/en/stable/) searches for and removes remnant adapter sequences from High-Throughput Sequencing (HTS) data and (optionally) trims low quality bases from the 3' end of reads following adapter removal. It is popular in the field of palaeogenomics. The output logs are stored in the results folder, and as a part of the MultiQC report.
diff --git a/workflows/mag.nf b/workflows/mag.nf index c27c7384..2f05e923 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -252,11 +252,9 @@ workflow MAG { ch_adapterremoval_pe_out = Channel.empty() ch_adapterremoval_pe_out = ADAPTERREMOVAL_PE.out.pair1_truncated .join(ADAPTERREMOVAL_PE.out.pair2_truncated) - .dump(tag: "ar_pe_mix_out") .map { [ it[0], [it[1], it[2]] ] } - .dump(tag: "ar2_pe_out") ch_short_reads = Channel.empty() ch_short_reads = ch_short_reads.mix(ADAPTERREMOVAL_SE.out.singles_truncated, ch_adapterremoval_pe_out) From ac21f02ecfaba98cbd314730f58d2e288a5665ed Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 12:14:14 +0100 Subject: [PATCH 124/246] prettier --- .github/workflows/ci.yml | 3 +- assets/multiqc_config.yml | 77 +++++++++++++++++++-------------------- docs/output.md | 4 +- nextflow_schema.json | 7 +--- 4 files changed, 44 insertions(+), 47 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 562d0d3e..dfa368a4 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -54,7 +54,8 @@ jobs: strategy: matrix: # Run remaining test profiles with minimum nextflow version - profile: [test_host_rm, test_hybrid, test_hybrid_host_rm, test_busco_auto, test_ancient_dna, test_adapterremoval] + profile: + [test_host_rm, test_hybrid, test_hybrid_host_rm, test_busco_auto, test_ancient_dna, test_adapterremoval] steps: - name: Check out pipeline code uses: actions/checkout@v2 diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 65639f09..3b5da468 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,52 +1,51 @@ report_comment: > - This report has been generated by the nf-core/mag - analysis pipeline. For information about how to interpret these results, please see the - documentation. + This report has been generated by the nf-core/mag + analysis pipeline. For information about how to interpret these results, please see the + documentation. report_section_order: - software_versions: - order: -1000 - nf-core-mag-summary: - order: -1001 + software_versions: + order: -1000 + nf-core-mag-summary: + order: -1001 export_plots: true -data_format: 'yaml' +data_format: "yaml" top_modules: -- 'fastqc': - name: 'FastQC: raw reads' - path_filters_exclude: - - '*trimmed*' -- 'fastp' -- 'adapterRemoval': -- custom_content -- 'fastqc': - name: 'FastQC: after preprocessing' - info: 'After trimming and, if requested, contamination removal.' - path_filters: - - '*trimmed*' -- 'busco': - info: 'assesses genome assembly and annotation completeness with Benchmarking Universal Single-Copy Orthologs. In case BUSCO''s automated lineage selection was used, only generic results for the selected domain are shown and only for genome bins and kept, unbinned contigs for which the BUSCO analysis was successfull, i.e. not for contigs for which no BUSCO genes could be found. Bins for which a specific virus lineage was selected are also not shown.' -- 'quast' - + - "fastqc": + name: "FastQC: raw reads" + path_filters_exclude: + - "*trimmed*" + - "fastp" + - "adapterRemoval": + - custom_content + - "fastqc": + name: "FastQC: after preprocessing" + info: "After trimming and, if requested, contamination removal." + path_filters: + - "*trimmed*" + - "busco": + info: "assesses genome assembly and annotation completeness with Benchmarking Universal Single-Copy Orthologs. In case BUSCO's automated lineage selection was used, only generic results for the selected domain are shown and only for genome bins and kept, unbinned contigs for which the BUSCO analysis was successfull, i.e. not for contigs for which no BUSCO genes could be found. Bins for which a specific virus lineage was selected are also not shown." + - "quast" custom_data: - host_removal: - file_format: 'tsv' - section_name: 'Bowtie 2: host read removal' - description: 'Reads are mapped against the host reference sequence. Only reads that do not align (concordantly) are kept for further analysis.' - plot_type: 'bargraph' - pconfig: - id: 'host_removal_bargraph' - title: 'Bowtie 2: reads mapped against host reference' - ylab: '# Reads' + host_removal: + file_format: "tsv" + section_name: "Bowtie 2: host read removal" + description: "Reads are mapped against the host reference sequence. Only reads that do not align (concordantly) are kept for further analysis." + plot_type: "bargraph" + pconfig: + id: "host_removal_bargraph" + title: "Bowtie 2: reads mapped against host reference" + ylab: "# Reads" sp: - host_removal: - fn: 'host_removal_metrics.tsv' - adapterRemoval: - fn: '*_ar2_*.log' + host_removal: + fn: "host_removal_metrics.tsv" + adapterRemoval: + fn: "*_ar2_*.log" extra_fn_clean_exts: - - '.bowtie2' - - '_ar2' + - ".bowtie2" + - "_ar2" diff --git a/docs/output.md b/docs/output.md index a667e5f7..e0b0fd89 100644 --- a/docs/output.md +++ b/docs/output.md @@ -64,8 +64,8 @@ FastQC is run for visualising the general quality metrics of the sequencing runs
Output files -* `QC_shortreads/adapterremoval/[sample]/` - * `[sample]_ar2.log`: AdapterRemoval log file (normally called `.settings` by AdapterRemoval.) +- `QC_shortreads/adapterremoval/[sample]/` + - `[sample]_ar2.log`: AdapterRemoval log file (normally called `.settings` by AdapterRemoval.)
diff --git a/nextflow_schema.json b/nextflow_schema.json index c405e935..b651e95a 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -278,10 +278,7 @@ "type": "string", "default": "fastp", "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", - "enum": [ - "fastp", - "adapterremoval" - ] + "enum": ["fastp", "adapterremoval"] }, "reads_minlength": { "type": "integer", @@ -703,4 +700,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} \ No newline at end of file +} From 087885d3713c5cf14ccf44757875235191ccc8c8 Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 12:19:40 +0100 Subject: [PATCH 125/246] add publish_dir_mode everywhere --- conf/modules.config | 15 +++++++++++---- 1 file changed, 11 insertions(+), 4 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 3a343302..ebbb4e3c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -53,7 +53,7 @@ process { ].join(' ').trim() publishDir = [ path: { "${params.outdir}/QC_shortreads/adapterremoval/${meta.id}" }, - mode: 'copy', + mode: params.publish_dir_mode, pattern: "*.{log}" ] ext.prefix = { "${meta.id}_ar2" } @@ -68,7 +68,7 @@ process { ].join(' ').trim() publishDir = [ path: { "${params.outdir}/QC_shortreads/adapterremoval/${meta.id}" }, - mode: 'copy', + mode: params.publish_dir_mode, pattern: "*.{log}" ] ext.prefix = { "${meta.id}_ar2" } @@ -307,6 +307,7 @@ process { ext.args = "-p ${params.freebayes_ploidy} -q ${params.freebayes_min_basequality} -F ${params.freebayes_minallelefreq}" publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/freebayes" }, + mode: params.publish_dir_mode, pattern: "*.vcf.gz" ] } @@ -316,6 +317,7 @@ process { ext.prefix = { "${meta.id}.filtered" } publishDir = [ path: { "${params.outdir}/Ancient_DNA/variant_calling/filtered" }, + mode: params.publish_dir_mode, pattern: "*.vcf.gz" ] } @@ -323,6 +325,7 @@ process { withName: BCFTOOLS_CONSENSUS { publishDir = [ path: {"${params.outdir}/Ancient_DNA/variant_calling/consensus" }, + mode: params.publish_dir_mode, pattern: "*.fa" ] } @@ -331,26 +334,30 @@ process { ext.args = "-t" publishDir = [ path: {"${params.outdir}/Ancient_DNA/variant_calling/index" }, + mode: params.publish_dir_mode, enabled: false ] } withName: PYDAMAGE_ANALYZE { publishDir = [ - path: {"${params.outdir}/Ancient_DNA/pydamage/analyze" } + path: {"${params.outdir}/Ancient_DNA/pydamage/analyze" }, + mode: params.publish_dir_mode ] } withName: PYDAMAGE_FILTER { ext.args = "-t ${params.pydamage_accuracy}" publishDir = [ - path: {"${params.outdir}/Ancient_DNA/pydamage/filter" } + path: {"${params.outdir}/Ancient_DNA/pydamage/filter" }, + mode: params.publish_dir_mode ] } withName: SAMTOOLS_FAIDX { publishDir = [ path: {"${params.outdir}/Ancient_DNA/samtools/faidx" }, + mode: params.publish_dir_mode, enabled: false ] } From 2e5470068dce48562d76fba2d162ca7025c4665e Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Thu, 24 Mar 2022 11:40:39 +0000 Subject: [PATCH 126/246] Template update for nf-core/tools version 2.3.2 --- .github/PULL_REQUEST_TEMPLATE.md | 1 - .github/workflows/awsfulltest.yml | 2 -- .github/workflows/awstest.yml | 2 -- .gitpod.yml | 2 +- CHANGELOG.md | 2 -- 5 files changed, 1 insertion(+), 8 deletions(-) diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 1e68020b..89c37cb4 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -10,7 +10,6 @@ Remember that PRs should be made against the dev branch, unless you're preparing Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/mag/tree/master/.github/CONTRIBUTING.md) --> - ## PR checklist diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index b74d2047..414181d9 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -22,8 +22,6 @@ jobs: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} - revision: ${{ github.sha }} workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/mag/work-${{ github.sha }} parameters: | { diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index f4aff861..c277a5cb 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -17,8 +17,6 @@ jobs: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} - revision: ${{ github.sha }} workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/mag/work-${{ github.sha }} parameters: | { diff --git a/.gitpod.yml b/.gitpod.yml index c452ee93..85d95ecc 100644 --- a/.gitpod.yml +++ b/.gitpod.yml @@ -4,7 +4,7 @@ vscode: extensions: # based on nf-core.nf-core-extensionpack - codezombiech.gitignore # Language support for .gitignore files # - cssho.vscode-svgviewer # SVG viewer - - davidanson.vscode-markdownlint # Markdown/CommonMark linting and style checking for Visual Studio Code + - esbenp.prettier-vscode # Markdown/CommonMark linting and style checking for Visual Studio Code - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar diff --git a/CHANGELOG.md b/CHANGELOG.md index 0f46b560..0803027f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,8 +11,6 @@ Initial release of nf-core/mag, created with the [nf-core](https://nf-co.re/) te ### `Fixed` -- Clarified conda usage and added an installation tutorial for Singularity since the one on Syllabs' website uses an outdate version of GO Compiler - ### `Dependencies` ### `Deprecated` From 4c5318fa9f03de164d2bc519338c8cec2502393f Mon Sep 17 00:00:00 2001 From: d4straub Date: Thu, 24 Mar 2022 13:16:48 +0100 Subject: [PATCH 127/246] update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 9f7923e5..f7a0f9e5 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,7 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Changed` -- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283) - Update to nf-core 2.3.1 `TEMPLATE` +- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289) - Update to nf-core 2.3.2 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint ### `Fixed` From ac8188ab6741ffb42502c9991b8684a8afda7a92 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 31 Mar 2022 14:48:31 +0200 Subject: [PATCH 128/246] Fix caching of metabat2 input --- subworkflows/local/binning_preparation.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/binning_preparation.nf b/subworkflows/local/binning_preparation.nf index e179e558..95e24da3 100644 --- a/subworkflows/local/binning_preparation.nf +++ b/subworkflows/local/binning_preparation.nf @@ -42,7 +42,7 @@ workflow BINNING_PREPARATION { // group mappings for one assembly ch_grouped_mappings = BOWTIE2_ASSEMBLY_ALIGN.out.mappings .groupTuple(by: 0) - .map { meta, assembly, bams, bais -> [ meta, assembly[0], bams, bais ] } // multiple symlinks to the same assembly -> use first + .map { meta, assembly, bams, bais -> [ meta, assembly.sort()[0], bams, bais ] } // multiple symlinks to the same assembly -> use first of sorted list emit: bowtie2_assembly_multiqc = BOWTIE2_ASSEMBLY_ALIGN.out.log.map { assembly_meta, reads_meta, log -> if (assembly_meta.id == reads_meta.id) {return [ log ]} } From 9b41688efc03a95c3cc86b16c5abaa6ab1b496ec Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 31 Mar 2022 17:28:18 +0200 Subject: [PATCH 129/246] Update CHANGELOG --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index f7a0f9e5..5d280292 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -19,6 +19,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Fixed` +- [#290](https://github.com/nf-core/mag/pull/290) - Fix caching of binning input + ### `Dependencies` | Tool | Previous version | New version | From 3e913835932112b533bdd22d14c9f3500522b784 Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 4 Apr 2022 15:09:14 +0200 Subject: [PATCH 130/246] update pydamage to v0.70 --- modules.json | 6 +++--- modules/nf-core/modules/pydamage/analyze/main.nf | 6 +++--- modules/nf-core/modules/pydamage/filter/main.nf | 6 +++--- 3 files changed, 9 insertions(+), 9 deletions(-) diff --git a/modules.json b/modules.json index bd8a90a2..4b91b0e9 100644 --- a/modules.json +++ b/modules.json @@ -46,14 +46,14 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "pydamage/analyze": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + "git_sha": "64b06baa06bc41269282bc7d286af37e859ad244" }, "pydamage/filter": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + "git_sha": "64b06baa06bc41269282bc7d286af37e859ad244" }, "samtools/faidx": { "git_sha": "1ad73f1b2abdea9398680d6d20014838135c9a35" } } } -} +} \ No newline at end of file diff --git a/modules/nf-core/modules/pydamage/analyze/main.nf b/modules/nf-core/modules/pydamage/analyze/main.nf index 39e01fa1..3463b0e5 100644 --- a/modules/nf-core/modules/pydamage/analyze/main.nf +++ b/modules/nf-core/modules/pydamage/analyze/main.nf @@ -2,10 +2,10 @@ process PYDAMAGE_ANALYZE { tag "$meta.id" label 'process_medium' - conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) + conda (params.enable_conda ? "bioconda::pydamage=0.70" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0' : - 'quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/pydamage:0.70--pyhdfd78af_0' : + 'quay.io/biocontainers/pydamage:0.70--pyhdfd78af_0' }" input: tuple val(meta), path(bam), path(bai) diff --git a/modules/nf-core/modules/pydamage/filter/main.nf b/modules/nf-core/modules/pydamage/filter/main.nf index a748875a..14fbf1c5 100644 --- a/modules/nf-core/modules/pydamage/filter/main.nf +++ b/modules/nf-core/modules/pydamage/filter/main.nf @@ -2,10 +2,10 @@ process PYDAMAGE_FILTER { tag "$meta.id" label 'process_low' - conda (params.enable_conda ? "bioconda::pydamage=0.62" : null) + conda (params.enable_conda ? "bioconda::pydamage=0.70" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/pydamage:0.62--pyhdfd78af_0' : - 'quay.io/biocontainers/pydamage:0.62--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/pydamage:0.70--pyhdfd78af_0' : + 'quay.io/biocontainers/pydamage:0.70--pyhdfd78af_0' }" input: tuple val(meta), path(csv) From a7c16f82b72fb7cd472d1faa007adeacc8724ea0 Mon Sep 17 00:00:00 2001 From: maxibor Date: Mon, 4 Apr 2022 15:17:48 +0200 Subject: [PATCH 131/246] update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 5d280292..d3f5b644 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -27,6 +27,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 | -------- | ---------------- | ----------- | | bcftools | 1.13 | 1.14 | | samtools | 1.14 | 1.15 | +| pydamage | 0.62 | 0.70 | ## v2.1.1 - 2021/11/25 From 577f2248073529e4bdb9bf81d7b700aa6ed8e793 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 31 Mar 2022 18:32:17 +0200 Subject: [PATCH 132/246] Change variable names --- subworkflows/local/binning.nf | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 6ba4b02d..81f69aa0 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -57,26 +57,26 @@ workflow BINNING { // combine depths back with assemblies ch_metabat2_input = assemblies - .map { meta, contigs, reads, indicies -> + .map { meta, assembly, bams, bais -> def meta_new = meta.clone() meta_new['binner'] = 'MetaBAT2' - [ meta_new, contigs, reads, indicies ] + [ meta_new, assembly, bams, bais ] } .join( ch_metabat_depths, by: 0 ) - .map { meta, contigs, reads, indicies, depths -> - [ meta, contigs, depths ] + .map { meta, assembly, bams, bais, depths -> + [ meta, assembly, depths ] } // conver metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) CONVERT_DEPTHS.out.output - .map { meta, contigs, reads, depth -> + .map { meta, assembly, reads, depth -> def meta_new = meta.clone() meta_new['binner'] = 'MaxBin2' - [ meta_new, contigs, reads, depth ] + [ meta_new, assembly, reads, depth ] } .set { ch_maxbin2_input } } From 84d7084d3018b99520c6f7afb82fd7c63d4d189a Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 8 Apr 2022 11:23:12 +0200 Subject: [PATCH 133/246] Run prettier --- modules.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules.json b/modules.json index 4b91b0e9..95a11bfc 100644 --- a/modules.json +++ b/modules.json @@ -56,4 +56,4 @@ } } } -} \ No newline at end of file +} From 77ba8d5fdfecc34405103988d079ca329ae45b31 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 29 Apr 2022 10:46:13 +0200 Subject: [PATCH 134/246] Code cleanup: replace .set{} with `=` everywhere for consistency and readabiltiy --- subworkflows/local/ancient_dna.nf | 3 +- subworkflows/local/binning.nf | 30 +++++-------- subworkflows/local/gtdbtk.nf | 6 +-- subworkflows/local/input_check.nf | 13 +++--- workflows/mag.nf | 73 ++++++++++--------------------- 5 files changed, 43 insertions(+), 82 deletions(-) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index c59c02e8..fae88f82 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -13,8 +13,7 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) FAIDX(input.map { item -> [ item[0], item[1] ] }) - input.join(FAIDX.out.fai) - .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] + freebayes_input = input.join(FAIDX.out.fai) // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], [], []] }, freebayes_input.map { item -> item[1] }, freebayes_input.map { item -> item[4] }, diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 81f69aa0..cd31f757 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -44,14 +44,13 @@ workflow BINNING { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) - METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth + ch_metabat_depths = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth .map { meta, depths -> def meta_new = meta.clone() meta_new['binner'] = 'MetaBAT2' [ meta_new, depths ] } - .set { ch_metabat_depths } ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions) @@ -71,14 +70,13 @@ workflow BINNING { // conver metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) - CONVERT_DEPTHS.out.output + ch_maxbin2_input = CONVERT_DEPTHS.out.output .map { meta, assembly, reads, depth -> def meta_new = meta.clone() meta_new['binner'] = 'MaxBin2' [ meta_new, assembly, reads, depth ] } - .set { ch_maxbin2_input } } // run binning @@ -106,31 +104,28 @@ workflow BINNING { // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for // MAG_DEPTHS, first have to separate and re-group due to limitation of // GUNZIP module - METABAT2_METABAT2.out.fasta.transpose().set { ch_metabat2_results_transposed } - MAXBIN2.out.binned_fastas.transpose().set { ch_maxbin2_results_transposed } + ch_metabat2_results_transposed = METABAT2_METABAT2.out.fasta.transpose() + ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() - SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } + ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) // mix all bins together - ch_metabat2_results_transposed + ch_final_bins_for_gunzip = ch_metabat2_results_transposed .mix( ch_maxbin2_results_transposed ) - .set { ch_final_bins_for_gunzip } GUNZIP_BINS ( ch_final_bins_for_gunzip ) - GUNZIP_BINS.out.gunzip - .set{ ch_binning_results_gunzipped } + ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions) GUNZIP_UNBINS ( ch_split_fasta_results_transposed ) - GUNZIP_UNBINS.out.gunzip - .set{ ch_splitfasta_results_gunzipped } + ch_splitfasta_results_gunzipped = GUNZIP_UNBINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions) // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta for grouping to mix back with original depth // files, as required for MAG_DEPTHS - ch_binning_results_gunzipped + ch_depth_input = ch_binning_results_gunzipped .mix(ch_splitfasta_results_gunzipped ) .map { meta, results -> def meta_new = meta.clone() @@ -138,7 +133,6 @@ workflow BINNING { } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) - .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) @@ -158,13 +152,11 @@ workflow BINNING { ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped + ch_binning_results_gunzipped_final = ch_binning_results_gunzipped .groupTuple(by: 0) - .set{ ch_binning_results_gunzipped_final } - METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) + ch_binning_results_gzipped_final = METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) .groupTuple(by: 0) - .set{ ch_binning_results_gzipped_final } SPLIT_FASTA.out.unbinned diff --git a/subworkflows/local/gtdbtk.nf b/subworkflows/local/gtdbtk.nf index 2fe0e933..fd971a33 100644 --- a/subworkflows/local/gtdbtk.nf +++ b/subworkflows/local/gtdbtk.nf @@ -19,7 +19,7 @@ workflow GTDBTK { // Filter bins: classify only medium & high quality MAGs // Collect completness and contamination metrics from busco summary def bin_metrics = [:] - busco_summary + ch_busco_metrics = busco_summary .splitCsv(header: true, sep: '\t') .map { row -> def completeness = -1 @@ -36,10 +36,9 @@ workflow GTDBTK { if (duplicated != '') contamination = Double.parseDouble(duplicated) [row.'GenomeBin', completeness, contamination] } - .set { ch_busco_metrics } // Filter bins based on collected metrics: completeness, contamination - bins + ch_filtered_bins = bins .transpose() .map { meta, bin -> [bin.getName(), bin, meta]} .join(ch_busco_metrics, failOnDuplicate: true, failOnMismatch: true) @@ -50,7 +49,6 @@ workflow GTDBTK { discarded: (it[2] == -1 || it[2] < params.gtdbtk_min_completeness || it[3] == -1 || it[3] > params.gtdbtk_max_contamination) return [it[0], it[1]] } - .set { ch_filtered_bins } GTDBTK_DB_PREPARATION ( gtdb ) GTDBTK_CLASSIFY ( diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index 348f4a16..2e4af3de 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -10,7 +10,7 @@ workflow INPUT_CHECK { main: if(hasExtension(params.input, "csv")){ // extracts read files from samplesheet CSV and distribute into channels - Channel + ch_input_rows = Channel .from(file(params.input)) .splitCsv(header: true) .map { row -> @@ -30,9 +30,9 @@ workflow INPUT_CHECK { exit 1, "Input samplesheet contains row with ${row.size()} column(s). Expects 5." } } - .set { ch_input_rows } + // separate short and long reads - ch_input_rows + ch_raw_short_reads = ch_input_rows .map { id, group, sr1, sr2, lr -> def meta = [:] meta.id = id @@ -43,8 +43,7 @@ workflow INPUT_CHECK { else return [ meta, [ sr1, sr2 ] ] } - .set { ch_raw_short_reads } - ch_input_rows + ch_raw_long_reads = ch_input_rows .map { id, group, sr1, sr2, lr -> if (lr) { def meta = [:] @@ -53,9 +52,8 @@ workflow INPUT_CHECK { return [ meta, lr ] } } - .set { ch_raw_long_reads } } else { - Channel + ch_raw_short_reads = Channel .fromFilePairs(params.input, size: params.single_end ? 1 : 2) .ifEmpty { exit 1, "Cannot find any reads matching: ${params.input}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --single_end on the command line." } .map { row -> @@ -65,7 +63,6 @@ workflow INPUT_CHECK { meta.single_end = params.single_end return [ meta, row[1] ] } - .set { ch_raw_short_reads } ch_input_rows = Channel.empty() ch_raw_long_reads = Channel.empty() } diff --git a/workflows/mag.nf b/workflows/mag.nf index a1857bda..616ca573 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -117,78 +117,66 @@ include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modu if ( params.host_genome ) { host_fasta = params.genomes[params.host_genome].fasta ?: false - Channel + ch_host_fasta = Channel .value(file( "${host_fasta}" )) - .set { ch_host_fasta } - host_bowtie2index = params.genomes[params.host_genome].bowtie2 ?: false - Channel + ch_host_bowtie2index = Channel .value(file( "${host_bowtie2index}/*" )) - .set { ch_host_bowtie2index } } else if ( params.host_fasta ) { - Channel + ch_host_fasta = Channel .value(file( "${params.host_fasta}" )) - .set { ch_host_fasta } } else { ch_host_fasta = Channel.empty() } if(params.busco_reference){ - Channel + ch_busco_db_file = Channel .value(file( "${params.busco_reference}" )) - .set { ch_busco_db_file } } else { ch_busco_db_file = Channel.empty() } if (params.busco_download_path) { - Channel + ch_busco_download_folder = Channel .value(file( "${params.busco_download_path}" )) - .set { ch_busco_download_folder } } else { ch_busco_download_folder = Channel.empty() } if(params.centrifuge_db){ - Channel + ch_centrifuge_db_file = Channel .value(file( "${params.centrifuge_db}" )) - .set { ch_centrifuge_db_file } } else { ch_centrifuge_db_file = Channel.empty() } if(params.kraken2_db){ - Channel + ch_kraken2_db_file = Channel .value(file( "${params.kraken2_db}" )) - .set { ch_kraken2_db_file } } else { ch_kraken2_db_file = Channel.empty() } if(params.cat_db){ - Channel + ch_cat_db_file = Channel .value(file( "${params.cat_db}" )) - .set { ch_cat_db_file } } else { ch_cat_db_file = Channel.empty() } if(!params.keep_phix) { - Channel + ch_phix_db_file = Channel .value(file( "${params.phix_reference}" )) - .set { ch_phix_db_file } } if (!params.keep_lambda) { - Channel + ch_nanolyse_db = Channel .value(file( "${params.lambda_reference}" )) - .set { ch_nanolyse_db } } gtdb = params.skip_busco ? false : params.gtdb if (gtdb) { - Channel + ch_gtdb = Channel .value(file( "${gtdb}" )) - .set { ch_gtdb } } else { ch_gtdb = Channel.empty() } @@ -325,15 +313,13 @@ workflow MAG { } // join long and short reads by sample name - ch_short_reads + ch_short_reads_tmp = ch_short_reads .map { meta, sr -> [ meta.id, meta, sr ] } - .set {ch_short_reads_tmp} - ch_long_reads + ch_short_and_long_reads = ch_long_reads .map { meta, lr -> [ meta.id, meta, lr ] } .join(ch_short_reads_tmp, by: 0) .map { id, meta_lr, lr, meta_sr, sr -> [ meta_lr, lr, sr[0], sr[1] ] } // should not occur for single-end, since SPAdes (hybrid) does not support single-end - .set{ ch_short_and_long_reads } FILTLONG ( ch_short_and_long_reads @@ -368,13 +354,12 @@ workflow MAG { if (( params.centrifuge_db || params.kraken2_db ) && !params.skip_krona){ KRONA_DB () - CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) + ch_tax_classifications = CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) . map { classifier, meta, report -> def meta_new = meta.clone() meta_new.classifier = classifier [ meta_new, report ] } - .set { ch_tax_classifications } KRONA ( ch_tax_classifications, KRONA_DB.out.db.collect() @@ -392,7 +377,7 @@ workflow MAG { if (params.coassemble_group) { // short reads // group and set group as new id - ch_short_reads + ch_short_reads_grouped = ch_short_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -403,10 +388,9 @@ workflow MAG { if (!params.single_end) [ meta, reads.collect { it[0] }, reads.collect { it[1] } ] else [ meta, reads.collect { it }, [] ] } - .set { ch_short_reads_grouped } // long reads // group and set group as new id - ch_long_reads + ch_long_reads_grouped = ch_long_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -415,25 +399,22 @@ workflow MAG { meta.group = group [ meta, reads.collect { it } ] } - .set { ch_long_reads_grouped } } else { - ch_short_reads + ch_short_reads_grouped = ch_short_reads .map { meta, reads -> if (!params.single_end){ [ meta, [reads[0]], [reads[1]] ] } else [ meta, [reads], [] ] } - .set { ch_short_reads_grouped } } ch_assemblies = Channel.empty() if (!params.skip_megahit){ MEGAHIT ( ch_short_reads_grouped ) - MEGAHIT.out.assembly + ch_megahit_assemblies = MEGAHIT.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "MEGAHIT" [ meta_new, assembly ] } - .set { ch_megahit_assemblies } ch_assemblies = ch_assemblies.mix(ch_megahit_assemblies) ch_versions = ch_versions.mix(MEGAHIT.out.versions.first()) } @@ -457,41 +438,36 @@ workflow MAG { } } else { ch_short_reads_spades = ch_short_reads - ch_long_reads + ch_long_reads_spades = ch_long_reads .map { meta, reads -> [ meta, [reads] ] } - .set { ch_long_reads_spades } } if (!params.single_end && !params.skip_spades){ SPADES ( ch_short_reads_spades ) - SPADES.out.assembly + ch_spades_assemblies = SPADES.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdes" [ meta_new, assembly ] } - .set { ch_spades_assemblies } ch_assemblies = ch_assemblies.mix(ch_spades_assemblies) ch_versions = ch_versions.mix(SPADES.out.versions.first()) } if (!params.single_end && !params.skip_spadeshybrid){ - ch_short_reads_spades + ch_short_reads_spades_tmp = ch_short_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } - .set {ch_short_reads_spades_tmp} - ch_long_reads_spades + ch_reads_spadeshybrid = ch_long_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } .combine(ch_short_reads_spades_tmp, by: 0) .map { id, meta_long, long_reads, meta_short, short_reads -> [ meta_short, long_reads, short_reads ] } - .set { ch_reads_spadeshybrid } SPADESHYBRID ( ch_reads_spadeshybrid ) - SPADESHYBRID.out.assembly + ch_spadeshybrid_assemblies = SPADESHYBRID.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdesHybrid" [ meta_new, assembly ] } - .set { ch_spadeshybrid_assemblies } ch_assemblies = ch_assemblies.mix(ch_spadeshybrid_assemblies) ch_versions = ch_versions.mix(SPADESHYBRID.out.versions.first()) } @@ -651,13 +627,12 @@ workflow MAG { /* * Prokka: Genome annotation */ - BINNING.out.bins.mix(BINNING.out.unbinned).transpose() + ch_bins_for_prokka = BINNING.out.bins.mix(BINNING.out.unbinned).transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() [ meta_new, bin ] } - .set { ch_bins_for_prokka } if (!params.skip_prokka){ PROKKA ( From 1897e9e411c8e35d1099c3da6d072b5aaec352a4 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 08:20:00 +0200 Subject: [PATCH 135/246] Simplify code snippet to remove meta info --- subworkflows/local/binning.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index cd31f757..2da81b75 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -129,7 +129,8 @@ workflow BINNING { .mix(ch_splitfasta_results_gunzipped ) .map { meta, results -> def meta_new = meta.clone() - [ [ 'id': meta_new['id'], 'group': meta_new['group'], 'single_end': meta_new['single_end'], 'assembler': meta_new['assembler'] ], results ] + meta_new.remove('binner') + [ meta_new, results ] } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) From fa557bb510904bbe6a9377736122538440719d49 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 10:07:27 +0200 Subject: [PATCH 136/246] Fix: separate bin depth files and heatmap plotting for different binners --- bin/get_mag_depths.py | 21 ++++++++++++++------- bin/get_mag_depths_summary.py | 4 ++-- bin/plot_mag_depths.py | 2 +- modules/local/mag_depths.nf | 8 ++++---- modules/local/mag_depths_plot.nf | 8 ++++---- subworkflows/local/binning.nf | 8 +++++++- 6 files changed, 32 insertions(+), 19 deletions(-) diff --git a/bin/get_mag_depths.py b/bin/get_mag_depths.py index 6cc547dd..9a13f591 100755 --- a/bin/get_mag_depths.py +++ b/bin/get_mag_depths.py @@ -15,8 +15,8 @@ def parse_args(args=None): parser = argparse.ArgumentParser() parser.add_argument('-b', '--bins' , required=True, nargs="+", metavar='FILE' , help="Bins: FASTA containing all contigs.") parser.add_argument('-d', '--depths' , required=True , metavar='FILE' , help="(Compressed) TSV file containing contig depths for each sample: contigName, contigLen, totalAvgDepth, sample1_avgDepth, sample1_var [, sample2_avgDepth, sample2_var, ...].") - parser.add_argument('-a', '--assembly_name', required=True , type=str , help="Assembly name.") - parser.add_argument('-o', "--out" , required=True , metavar='FILE', type=argparse.FileType('w'), help="Output file containing depth for each bin.") + parser.add_argument('-a', '--assembler' , required=True , type=str , help="Assembler name.") + parser.add_argument('-i', '--id' , required=True , type=str , help="Sample or group id.") return parser.parse_args(args) def main(args=None): @@ -31,8 +31,8 @@ def main(args=None): header = next(reader) for sample in range(int((len(header)-3)/2)): col_name = header[3+2*sample] - # retrieve sample name: "-.bam" - sample_name = col_name[len(args.assembly_name)+1:-4] + # retrieve sample name: "--.bam" + sample_name = col_name[len(args.assembler)+1+len(args.id)+1:-4] sample_names.append(sample_name) # process contig depths for row in reader: @@ -41,10 +41,14 @@ def main(args=None): contig_depths.append(float(row[3+2*sample])) dict_contig_depths[str(row[0])] = contig_depths + # Initialize output files n_samples = len(sample_names) - # for each bin, access contig depths and compute mean bin depth (for all samples) - print("bin", '\t'.join(sample_names), sep='\t', file=args.out) + binners = set([ os.path.basename(file).split("-")[1] for file in args.bins ]) + for binner in binners: + with open(args.assembler + "-" + binner + "-" + args.id + "-binDepths.tsv", 'w') as outfile: + print("bin", '\t'.join(sample_names), sep='\t', file=outfile) + # for each bin, access contig depths and compute mean bin depth (for all samples) for file in args.bins: all_depths = [[] for i in range(n_samples)] @@ -60,7 +64,10 @@ def main(args=None): contig_depths = dict_contig_depths[rec.id] for sample in range(n_samples): all_depths[sample].append(contig_depths[sample]) - print(os.path.basename(file), '\t'.join(str(statistics.median(sample_depths)) for sample_depths in all_depths), sep='\t', file=args.out) + + binname = os.path.basename(file) + with open(args.assembler + "-" + binname.split("-")[1] + "-" + args.id + "-binDepths.tsv", 'a') as outfile: + print(binname, '\t'.join(str(statistics.median(sample_depths)) for sample_depths in all_depths), sep='\t', file=outfile) if __name__ == "__main__": diff --git a/bin/get_mag_depths_summary.py b/bin/get_mag_depths_summary.py index da5207d4..e70e640e 100755 --- a/bin/get_mag_depths_summary.py +++ b/bin/get_mag_depths_summary.py @@ -7,8 +7,8 @@ def parse_args(args=None): parser = argparse.ArgumentParser() - parser.add_argument('-d', '--depths' , required=True, nargs="+", metavar='FILE' , help="TSV file for each assembly containing bin depths for samples: bin, sample1, ....") - parser.add_argument('-o', "--out" , required=True , metavar='FILE', type=argparse.FileType('w'), help="Output file containing depths for all assemblies and all samples.") + parser.add_argument('-d', '--depths' , required=True, nargs="+", metavar='FILE' , help="TSV file for each assembly and binning method containing bin depths for samples: bin, sample1, ....") + parser.add_argument('-o', "--out" , required=True , metavar='FILE', type=argparse.FileType('w'), help="Output file containing depths for all assemblies, binning methods and all samples.") return parser.parse_args(args) def main(args=None): diff --git a/bin/plot_mag_depths.py b/bin/plot_mag_depths.py index 20de079c..5e7bff24 100755 --- a/bin/plot_mag_depths.py +++ b/bin/plot_mag_depths.py @@ -11,7 +11,7 @@ def parse_args(args=None): parser = argparse.ArgumentParser() - parser.add_argument('-d', '--bin_depths' , required=True, metavar='FILE' , help="Bin depths file in TSV format (for one assembly): bin, sample1_depth, sample2_depth, ....") + parser.add_argument('-d', '--bin_depths' , required=True, metavar='FILE' , help="Bin depths file in TSV format (for one assembly and binning method): bin, sample1_depth, sample2_depth, ....") parser.add_argument('-g', '--groups' , required=True, metavar='FILE' , help="File in TSV format containing group information for samples: sample, group") parser.add_argument('-o', "--out" , required=True, metavar='FILE', type=str, help="Output file.") return parser.parse_args(args) diff --git a/modules/local/mag_depths.nf b/modules/local/mag_depths.nf index f013492a..b7e09f23 100644 --- a/modules/local/mag_depths.nf +++ b/modules/local/mag_depths.nf @@ -11,15 +11,15 @@ process MAG_DEPTHS { tuple val(meta), path(bins), path(contig_depths) output: - tuple val(meta), path("${meta.assembler}-${meta.id}-binDepths.tsv"), emit: depths - path "versions.yml" , emit: versions + tuple val(meta), path("${meta.assembler}-*-${meta.id}-binDepths.tsv"), emit: depths + path "versions.yml" , emit: versions script: """ get_mag_depths.py --bins ${bins} \\ --depths ${contig_depths} \\ - --assembly_name "${meta.assembler}-${meta.id}" \\ - --out "${meta.assembler}-${meta.id}-binDepths.tsv" + --assembler ${meta.assembler} \\ + --id ${meta.id} cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/mag_depths_plot.nf b/modules/local/mag_depths_plot.nf index 16435d46..9a6b61e7 100644 --- a/modules/local/mag_depths_plot.nf +++ b/modules/local/mag_depths_plot.nf @@ -1,5 +1,5 @@ process MAG_DEPTHS_PLOT { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" conda (params.enable_conda ? "conda-forge::python=3.9 conda-forge::pandas=1.3.0 anaconda::seaborn=0.11.0" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -11,14 +11,14 @@ process MAG_DEPTHS_PLOT { path(sample_groups) output: - tuple val(meta), path("${meta.assembler}-${meta.id}-binDepths.heatmap.png"), emit: heatmap - path "versions.yml" , emit: versions + tuple val(meta), path("${meta.assembler}-${meta.binner}-${meta.id}-binDepths.heatmap.png"), emit: heatmap + path "versions.yml" , emit: versions script: """ plot_mag_depths.py --bin_depths ${depths} \ --groups ${sample_groups} \ - --out "${meta.assembler}-${meta.id}-binDepths.heatmap.png" + --out "${meta.assembler}-${meta.binner}-${meta.id}-binDepths.heatmap.png" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 2da81b75..7c57c7ae 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -143,9 +143,15 @@ workflow BINNING { ch_sample_groups = reads .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } + // Transpose and add 'binner' meta information again for plotting // filter MAG depth files: use only those for plotting that contain depths for > 2 samples ch_mag_depths_plot = MAG_DEPTHS.out.depths - .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } + .transpose() + .map { meta, depth_file -> + def meta_new = meta.clone() + meta_new['binner'] = depth_file.name.split("-")[1] + if (getColNo(depth_file) > 2) [ meta_new, depth_file ] + } MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) From f7d9fa45fc152127cec28eb0314269e2adb00175 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 10:10:43 +0200 Subject: [PATCH 137/246] Update docs for binner separated heatmaps --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index e0b0fd89..94ddefae 100644 --- a/docs/output.md +++ b/docs/output.md @@ -336,7 +336,7 @@ For each genome bin the median sequencing depth is computed based on the corresp - `GenomeBinning/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. + - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`.
From dc0c5bdb39add47c42d8cf5d77e8fa2f1114f11d Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 10:41:25 +0200 Subject: [PATCH 138/246] Add extra results folder for depth files --- conf/modules.config | 12 ++++++++++-- 1 file changed, 10 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index ebbb4e3c..650e95d7 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -201,7 +201,15 @@ process { ] } - withName: 'MAG_DEPTHS_PLOT|MAG_DEPTHS_SUMMARY|BIN_SUMMARY' { + withName: 'MAG_DEPTHS_PLOT|MAG_DEPTHS_SUMMARY' { + publishDir = [ + path: { "${params.outdir}/GenomeBinning/depths" }, + mode: params.publish_dir_mode, + pattern: "*.{png,tsv}" + ] + } + + withName: 'BIN_SUMMARY' { publishDir = [ path: { "${params.outdir}/GenomeBinning" }, mode: params.publish_dir_mode, @@ -363,7 +371,7 @@ process { } withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { publishDir = [ - path: { "${params.outdir}/GenomeBinning/" }, + path: { "${params.outdir}/GenomeBinning/depths" }, mode: params.publish_dir_mode, pattern: '*-depth.txt.gz' ] From 88d46b060d160ee32ea70f523a5b29a0f8ed2202 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 10:43:46 +0200 Subject: [PATCH 139/246] Update docs for new depths results folder --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index 94ddefae..48b2abd5 100644 --- a/docs/output.md +++ b/docs/output.md @@ -261,7 +261,7 @@ Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig
Output files -- `GenomeBinning/` +- `GenomeBinning/depths/` - `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads.
@@ -334,7 +334,7 @@ For each genome bin the median sequencing depth is computed based on the corresp
Output files -- `GenomeBinning/` +- `GenomeBinning/depths/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. From 732b71b1190e3325dec72fee1eab4780fc672ea3 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 13 May 2022 11:31:45 +0200 Subject: [PATCH 140/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d3f5b644..be27e2d5 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -16,6 +16,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289) - Update to nf-core 2.3.2 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint +- [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths` for `[assembler]-[sample/group]-depth.txt.gz`, `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` ### `Fixed` From f28652bbd6db81c894ad0900a8da1c37045067c9 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 24 Mar 2022 17:22:00 +0100 Subject: [PATCH 141/246] Start work on DAS_Tool --- CITATIONS.md | 8 ++ modules.json | 8 +- .../nf-core/modules/dastool/dastool/main.nf | 67 +++++++++++++ .../nf-core/modules/dastool/dastool/meta.yml | 99 +++++++++++++++++++ .../modules/dastool/scaffolds2bin/main.nf | 40 ++++++++ .../modules/dastool/scaffolds2bin/meta.yml | 58 +++++++++++ nextflow.config | 5 +- subworkflows/local/binning_refinement.nf | 47 +++++++++ workflows/mag.nf | 11 +++ 9 files changed, 340 insertions(+), 3 deletions(-) create mode 100644 modules/nf-core/modules/dastool/dastool/main.nf create mode 100644 modules/nf-core/modules/dastool/dastool/meta.yml create mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/main.nf create mode 100644 modules/nf-core/modules/dastool/scaffolds2bin/meta.yml create mode 100644 subworkflows/local/binning_refinement.nf diff --git a/CITATIONS.md b/CITATIONS.md index 387ad6c3..8d9ac2ac 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,6 +10,10 @@ ## Pipeline tools +- [AdapterRemoval2](https://doi.org/10.1186/) + + > Schubert, M., Lindgreen, S., and Orlando, L. 2016. "AdapterRemoval v2: Rapid Adapter Trimming, Identification, and Read Merging." BMC Research Notes 9 (February): 88. doi: 10.1186/s13104-016-1900-2 + - [BCFtools](https://doi.org/10.1093/gigascience/giab008) > Danecek, Petr, et al. "Twelve years of SAMtools and BCFtools." Gigascience 10.2 (2021): giab008. doi: 10.1093/gigascience/giab008 @@ -30,6 +34,10 @@ > Kim, D., Song, L., Breitwieser, F. P., & Salzberg, S. L. (2016). Centrifuge: rapid and sensitive classification of metagenomic sequences. Genome research, 26(12), 1721-1729. doi: 10.1101/gr.210641.116. +- [DAS_Tool](https://doi.org/10.1038/s41564-018-0171-1) + + > Sieber, C. M. K., et al. 2018. "Recovery of Genomes from Metagenomes via a Dereplication, Aggregation and Scoring Strategy." Nature Microbiology 3 (7): 836-43. doi: 10.1038/s41564-018-0171-1 + - [FastP](https://doi.org/10.1093/bioinformatics/bty560) > Chen, S., Zhou, Y., Chen, Y., & Gu, J. (2018). fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics , 34(17), i884–i890. doi: 10.1093/bioinformatics/bty560. diff --git a/modules.json b/modules.json index 95a11bfc..3807b1f5 100644 --- a/modules.json +++ b/modules.json @@ -18,6 +18,12 @@ "custom/dumpsoftwareversions": { "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, + "dastool/dastool": { + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + }, + "dastool/scaffolds2bin": { + "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + }, "fastp": { "git_sha": "d0a1cbb703a130c19f6796c3fce24fbe7dfce789" }, @@ -56,4 +62,4 @@ } } } -} +} \ No newline at end of file diff --git a/modules/nf-core/modules/dastool/dastool/main.nf b/modules/nf-core/modules/dastool/dastool/main.nf new file mode 100644 index 00000000..53dfea19 --- /dev/null +++ b/modules/nf-core/modules/dastool/dastool/main.nf @@ -0,0 +1,67 @@ +process DASTOOL_DASTOOL { + tag "$meta.id" + label 'process_medium' + + conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : + 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + + input: + tuple val(meta), path(contigs), path(bins) + path(proteins) + path(db_directory) + val(search_engine) + + output: + tuple val(meta), path("*.log") , emit: log + tuple val(meta), path("*_summary.txt") , emit: summary + tuple val(meta), path("*_DASTool_scaffolds2bin.txt") , emit: scaffolds2bin + tuple val(meta), path("*.eval") , optional: true, emit: eval + tuple val(meta), path("*_DASTool_bins/*.fa") , optional: true, emit: bins + tuple val(meta), path("*.pdf") , optional: true, emit: pdfs + tuple val(meta), path("*.proteins.faa") , optional: true, emit: fasta_proteins + tuple val(meta), path("*.archaea.scg") , optional: true, emit: fasta_archaea_scg + tuple val(meta), path("*.bacteria.scg") , optional: true, emit: fasta_bacteria_scg + tuple val(meta), path("*.seqlength") , optional: true, emit: seqlength + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def bin_list = bins instanceof List ? bins.join(",") : "$bins" + def engine = search_engine ? "--search_engine $search_engine" : "--search_engine diamond" + def db_dir = db_directory ? "--db_directory $db_directory" : "" + def clean_contigs = contigs.toString() - ".gz" + def decompress_contigs = contigs.toString() == clean_contigs ? "" : "gunzip -q -f $contigs" + def decompress_proteins = proteins ? "gunzip -f $proteins" : "" + def clean_proteins = proteins ? proteins.toString() - ".gz" : "" + def proteins_pred = proteins ? "--proteins $clean_proteins" : "" + + if (! search_engine) { + log.info('[DAS_Tool] Default search engine (USEARCH) is proprietary software and not available in bioconda. Using DIAMOND as alternative.') + } + + """ + $decompress_proteins + $decompress_contigs + + DAS_Tool \\ + $args \\ + $proteins_pred \\ + $db_dir \\ + $engine \\ + -t $task.cpus \\ + --bins $bin_list \\ + -c $clean_contigs \\ + -o $prefix + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/dastool/dastool/meta.yml b/modules/nf-core/modules/dastool/dastool/meta.yml new file mode 100644 index 00000000..a77df9bd --- /dev/null +++ b/modules/nf-core/modules/dastool/dastool/meta.yml @@ -0,0 +1,99 @@ +name: dastool_dastool +description: DAS Tool binning step. +keywords: + - binning + - das tool + - table + - de novo + - bins + - contigs + - assembly + - das_tool +tools: + - dastool: + description: | + DAS Tool is an automated method that integrates the results + of a flexible number of binning algorithms to calculate an optimized, non-redundant + set of bins from a single assembly. + + homepage: https://github.com/cmks/DAS_Tool + documentation: https://github.com/cmks/DAS_Tool + tool_dev_url: https://github.com/cmks/DAS_Tool + doi: "10.1038/s41564-018-0171-1" + licence: ["BSD"] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - contigs: + type: file + description: fasta file + pattern: "*.{fa.gz,fas.gz,fasta.gz}" + - bins: + type: file + description: "Scaffolds2bin tabular file generated with dastool/scaffolds2bin" + pattern: "*.scaffolds2bin.tsv" + - proteins: + type: file + description: Predicted proteins in prodigal fasta format (>scaffoldID_geneNo) + pattern: "*.{fa.gz,fas.gz,fasta.gz}" + - db_directory: + type: file + description: (optional) Directory of single copy gene database. + - search_engine: + type: val + description: Engine used for single copy gene identification. USEARCH is not supported due to it being proprietary [blast/diamond] + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - version: + type: file + description: File containing software version + pattern: "versions.yml" + - log: + type: file + description: Log file of the run + pattern: "*.log" + - summary: + type: file + description: Summary of output bins including quality and completeness estimates + pattern: "*summary.txt" + - scaffolds2bin: + type: file + description: Scaffolds to bin file of output bins + pattern: "*.scaffolds2bin.txt" + - eval: + type: file + description: Quality and completeness estimates of input bin sets + pattern: "*.eval" + - pdfs: + type: file + description: Plots showing the amount of high quality bins and score distribution of bins per method + pattern: "*.pdf" + - fasta_proteins: + type: file + description: Output from prodigal if not already supplied + pattern: "*.proteins.faa" + - fasta_archaea_scg: + type: file + description: Results of archaeal single-copy-gene prediction + pattern: "*.archaea.scg" + - fasta_bacteria_scg: + type: file + description: Results of bacterial single-copy-gene prediction + pattern: "*.bacteria.scg" + - seqlength: + type: file + description: Summary of contig lengths + pattern: "*.seqlength" + +authors: + - "@maxibor" + - "@jfy133" diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf b/modules/nf-core/modules/dastool/scaffolds2bin/main.nf new file mode 100644 index 00000000..6e9fcdc2 --- /dev/null +++ b/modules/nf-core/modules/dastool/scaffolds2bin/main.nf @@ -0,0 +1,40 @@ +process DASTOOL_SCAFFOLDS2BIN { + tag "$meta.id" + label 'process_low' + + conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : + 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + + input: + tuple val(meta), path(fasta) + val(extension) + + output: + tuple val(meta), path("*.tsv"), emit: scaffolds2bin + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def file_extension = extension ? extension : "fasta" + + """ + gunzip -f *.${file_extension}.gz + + Fasta_to_Scaffolds2Bin.sh \\ + $args \\ + -i . \\ + -e $file_extension \\ + > ${prefix}.tsv + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml b/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml new file mode 100644 index 00000000..0bf8618d --- /dev/null +++ b/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml @@ -0,0 +1,58 @@ +name: dastool_scaffolds2bin +description: Helper script to convert a set of bins in fasta format to tabular scaffolds2bin format +keywords: + - binning + - das tool + - table + - de novo + - bins + - contigs + - assembly + - das_tool +tools: + - dastool: + description: | + DAS Tool is an automated method that integrates the results + of a flexible number of binning algorithms to calculate an optimized, non-redundant + set of bins from a single assembly. + + homepage: https://github.com/cmks/DAS_Tool + documentation: https://github.com/cmks/DAS_Tool + tool_dev_url: https://github.com/cmks/DAS_Tool + doi: "10.1038/s41564-018-0171-1" + licence: ["BSD"] + +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Fasta of list of fasta files recommended to be gathered via with .collect() of bins + pattern: "*.{fa,fas,fasta}" + - binner: + type: val + description: Name of the binning software (optional) + - extension: + type: val + description: Fasta file extension (fa | fas | fasta | ...) + +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - scaffolds2bin: + type: file + description: tabular scaffolds2bin file for DAS tool input + pattern: "*.scaffolds2bin.tsv" + +authors: + - "@maxibor" diff --git a/nextflow.config b/nextflow.config index 6dcef6d5..0583d448 100644 --- a/nextflow.config +++ b/nextflow.config @@ -85,8 +85,9 @@ params { lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // binning options - skip_metabat2 = false - skip_maxbin2 = false + skip_metabat2 = false + skip_maxbin2 = false + refine_bins_dastool = false // Bin QC skip_busco = false diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf new file mode 100644 index 00000000..f8d7ae9c --- /dev/null +++ b/subworkflows/local/binning_refinement.nf @@ -0,0 +1,47 @@ +/* + * Binning with MetaBAT2 and MaxBin2 + */ + +include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' + +workflow BINNING_REFINEMENT { + take: + contigs // + bins // channel: [ val(meta), path(bins) ] + + main: + ch_versions = Channel.empty() + + ch_contigs_for_dastool = contigs + .map { + meta, assembly, bams, bais -> + def meta_new = meta.clone() + [ meta_new, assembly ] + } + .dump(tag: "binrefine_contigs") + bins.dump(tag: "binrefine_bins") + + ch_bins_for_scaffolds2bin = bins + .branch { + metabat2: it[0]['binner'] == 'MetaBAT2' + maxbin2: it[0]['binner'] == 'MaxBin2' + } + + // Run on each bin file separately + DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_bins_for_scaffolds2bin.metabat2, "fa") + DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_bins_for_scaffolds2bin.maxbin2, "fasta") + + ch_versions = ch_versions.mix(DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions) + ch_versions = ch_versions.mix(DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions) + + // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) + + // Run DAStool + //DASTOOL_DASTOOL() + //ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions) + + emit: + versions = ch_versions +} diff --git a/workflows/mag.nf b/workflows/mag.nf index 616ca573..08d170c5 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -89,6 +89,7 @@ include { MULTIQC } from '../modules include { INPUT_CHECK } from '../subworkflows/local/input_check' include { BINNING_PREPARATION } from '../subworkflows/local/binning_preparation' include { BINNING } from '../subworkflows/local/binning' +include { BINNING_REFINEMENT } from '../subworkflows/local/binning_refinement' include { BUSCO_QC } from '../subworkflows/local/busco_qc' include { GTDBTK } from '../subworkflows/local/gtdbtk' include { ANCIENT_DNA_ASSEMLY_VALIDATION } from '../subworkflows/local/ancient_dna' @@ -642,8 +643,18 @@ workflow MAG { ) ch_versions = ch_versions.mix(PROKKA.out.versions.first()) } + + /* + * DAS_Tool: binning refinement + */ + if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { + BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) + } + } + + CUSTOM_DUMPSOFTWAREVERSIONS ( ch_versions.unique().collectFile(name: 'collated_versions.yml') ) From 7379e6463cc9add79c5a82622df041c07b50621b Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Sat, 2 Apr 2022 08:23:18 +0200 Subject: [PATCH 142/246] Update DASTool move test to hybrid --- conf/modules.config | 37 +++++++++++++++++++ conf/test.config | 16 ++++---- conf/test_ancient_dna.config | 2 + conf/test_hybrid.config | 2 + modules.json | 6 +-- .../nf-core/modules/dastool/dastool/main.nf | 29 ++++++--------- .../nf-core/modules/dastool/dastool/meta.yml | 18 +++++---- .../main.nf | 21 ++++++----- .../meta.yml | 16 ++++---- nextflow.config | 7 ++-- subworkflows/local/binning.nf | 18 ++++----- subworkflows/local/binning_refinement.nf | 37 ++++++++++++++----- workflows/mag.nf | 2 +- 13 files changed, 134 insertions(+), 77 deletions(-) rename modules/nf-core/modules/dastool/{scaffolds2bin => fastatocontig2bin}/main.nf (59%) rename modules/nf-core/modules/dastool/{scaffolds2bin => fastatocontig2bin}/meta.yml (83%) diff --git a/conf/modules.config b/conf/modules.config index 650e95d7..baeb97f4 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -443,6 +443,43 @@ process { ] } + withName: DASTOOL_FASTATOCONTIG2BIN_METABAT2 { + publishDir = [ + [ + path: { "${params.outdir}/BinRefinement/fastatocontig2bin" }, + mode: params.publish_dir_mode, + pattern: '*.tsv' + ], + ] + ext.prefix = { "${meta.assembler}-MetaBAT2-${meta.id}" } + + } + + withName: DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 { + publishDir = [ + [ + path: { "${params.outdir}/BinRefinement/fastatocontig2bin" }, + mode: params.publish_dir_mode, + pattern: '*.tsv' + ], + ] + ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } + + } + + withName: DASTOOL_DASTOOL { + publishDir = [ + [ + path: { "${params.outdir}/BinRefinement/" }, + mode: params.publish_dir_mode, + pattern: '*.tsv' + ], + ] + ext.prefix = { "${meta.assembler}-${meta.id}" } + // TODO move penalties to test config if they work! + ext.args = "--write_bins --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" + } + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/conf/test.config b/conf/test.config index 8b381128..5df32bdb 100644 --- a/conf/test.config +++ b/conf/test.config @@ -20,12 +20,12 @@ params { max_time = '6.h' // Input data - input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' - centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" - kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" - skip_krona = true - min_length_unbinned_contigs = 1 - max_unbinned_contigs = 2 - busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" - gtdb = false + input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' + centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" + kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" + skip_krona = true + min_length_unbinned_contigs = 1 + max_unbinned_contigs = 2 + busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" + gtdb = false } diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config index d4c621ef..84e18830 100644 --- a/conf/test_ancient_dna.config +++ b/conf/test_ancient_dna.config @@ -33,4 +33,6 @@ params { skip_spades = false skip_spadeshybrid = true bcftools_view_variant_quality = 0 + refine_bins_dastool = true + refine_bins_dastool_threshold = 0 } diff --git a/conf/test_hybrid.config b/conf/test_hybrid.config index edc879e7..6ba74bfb 100644 --- a/conf/test_hybrid.config +++ b/conf/test_hybrid.config @@ -25,4 +25,6 @@ params { max_unbinned_contigs = 2 busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" gtdb = false + refine_bins_dastool = true + refine_bins_dastool_threshold = 0 } diff --git a/modules.json b/modules.json index 3807b1f5..69f7927f 100644 --- a/modules.json +++ b/modules.json @@ -19,10 +19,10 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "dastool/dastool": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + "git_sha": "8ce68107871c96519b3eb0095d97896e34ef4489" }, - "dastool/scaffolds2bin": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + "dastool/fastatocontig2bin": { + "git_sha": "8ce68107871c96519b3eb0095d97896e34ef4489" }, "fastp": { "git_sha": "d0a1cbb703a130c19f6796c3fce24fbe7dfce789" diff --git a/modules/nf-core/modules/dastool/dastool/main.nf b/modules/nf-core/modules/dastool/dastool/main.nf index 53dfea19..968f85de 100644 --- a/modules/nf-core/modules/dastool/dastool/main.nf +++ b/modules/nf-core/modules/dastool/dastool/main.nf @@ -2,27 +2,28 @@ process DASTOOL_DASTOOL { tag "$meta.id" label 'process_medium' - conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + conda (params.enable_conda ? "bioconda::das_tool=1.1.4" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : - 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.4--r41hdfd78af_1' : + 'quay.io/biocontainers/das_tool:1.1.4--r41hdfd78af_1' }" input: tuple val(meta), path(contigs), path(bins) path(proteins) path(db_directory) - val(search_engine) output: tuple val(meta), path("*.log") , emit: log - tuple val(meta), path("*_summary.txt") , emit: summary - tuple val(meta), path("*_DASTool_scaffolds2bin.txt") , emit: scaffolds2bin + tuple val(meta), path("*_summary.tsv") , emit: summary + tuple val(meta), path("*_DASTool_contig2bin.tsv") , emit: contig2bin tuple val(meta), path("*.eval") , optional: true, emit: eval tuple val(meta), path("*_DASTool_bins/*.fa") , optional: true, emit: bins tuple val(meta), path("*.pdf") , optional: true, emit: pdfs - tuple val(meta), path("*.proteins.faa") , optional: true, emit: fasta_proteins + tuple val(meta), path("*.candidates.faa") , optional: true, emit: fasta_proteins + tuple val(meta), path("*.faa") , optional: true, emit: candidates_faa tuple val(meta), path("*.archaea.scg") , optional: true, emit: fasta_archaea_scg tuple val(meta), path("*.bacteria.scg") , optional: true, emit: fasta_bacteria_scg + tuple val(meta), path("*.b6") , optional: true, emit: b6 tuple val(meta), path("*.seqlength") , optional: true, emit: seqlength path "versions.yml" , emit: versions @@ -33,17 +34,12 @@ process DASTOOL_DASTOOL { def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" def bin_list = bins instanceof List ? bins.join(",") : "$bins" - def engine = search_engine ? "--search_engine $search_engine" : "--search_engine diamond" def db_dir = db_directory ? "--db_directory $db_directory" : "" def clean_contigs = contigs.toString() - ".gz" def decompress_contigs = contigs.toString() == clean_contigs ? "" : "gunzip -q -f $contigs" - def decompress_proteins = proteins ? "gunzip -f $proteins" : "" def clean_proteins = proteins ? proteins.toString() - ".gz" : "" - def proteins_pred = proteins ? "--proteins $clean_proteins" : "" - - if (! search_engine) { - log.info('[DAS_Tool] Default search engine (USEARCH) is proprietary software and not available in bioconda. Using DIAMOND as alternative.') - } + def decompress_proteins = proteins ? "gunzip -f $proteins" : "" + def proteins_pred = proteins ? "-p $clean_proteins" : "" """ $decompress_proteins @@ -53,15 +49,14 @@ process DASTOOL_DASTOOL { $args \\ $proteins_pred \\ $db_dir \\ - $engine \\ -t $task.cpus \\ - --bins $bin_list \\ + -i $bin_list \\ -c $clean_contigs \\ -o $prefix cat <<-END_VERSIONS > versions.yml "${task.process}": - dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool //' ) END_VERSIONS """ } diff --git a/modules/nf-core/modules/dastool/dastool/meta.yml b/modules/nf-core/modules/dastool/dastool/meta.yml index a77df9bd..0889ca47 100644 --- a/modules/nf-core/modules/dastool/dastool/meta.yml +++ b/modules/nf-core/modules/dastool/dastool/meta.yml @@ -34,8 +34,8 @@ input: pattern: "*.{fa.gz,fas.gz,fasta.gz}" - bins: type: file - description: "Scaffolds2bin tabular file generated with dastool/scaffolds2bin" - pattern: "*.scaffolds2bin.tsv" + description: "FastaToContig2Bin tabular file generated with dastool/fastatocontig2bin" + pattern: "*.tsv" - proteins: type: file description: Predicted proteins in prodigal fasta format (>scaffoldID_geneNo) @@ -43,9 +43,6 @@ input: - db_directory: type: file description: (optional) Directory of single copy gene database. - - search_engine: - type: val - description: Engine used for single copy gene identification. USEARCH is not supported due to it being proprietary [blast/diamond] output: - meta: @@ -65,14 +62,17 @@ output: type: file description: Summary of output bins including quality and completeness estimates pattern: "*summary.txt" - - scaffolds2bin: + - contig2bin: type: file description: Scaffolds to bin file of output bins - pattern: "*.scaffolds2bin.txt" + pattern: "*.contig2bin.txt" - eval: type: file description: Quality and completeness estimates of input bin sets pattern: "*.eval" + - bins: + description: Final refined bins in fasta format + pattern: "*.fa" - pdfs: type: file description: Plots showing the amount of high quality bins and score distribution of bins per method @@ -89,6 +89,10 @@ output: type: file description: Results of bacterial single-copy-gene prediction pattern: "*.bacteria.scg" + - b6: + type: file + description: Results in b6 format + pattern: "*.b6" - seqlength: type: file description: Summary of contig lengths diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf b/modules/nf-core/modules/dastool/fastatocontig2bin/main.nf similarity index 59% rename from modules/nf-core/modules/dastool/scaffolds2bin/main.nf rename to modules/nf-core/modules/dastool/fastatocontig2bin/main.nf index 6e9fcdc2..8bb13380 100644 --- a/modules/nf-core/modules/dastool/scaffolds2bin/main.nf +++ b/modules/nf-core/modules/dastool/fastatocontig2bin/main.nf @@ -1,19 +1,19 @@ -process DASTOOL_SCAFFOLDS2BIN { +process DASTOOL_FASTATOCONTIG2BIN { tag "$meta.id" label 'process_low' - conda (params.enable_conda ? "bioconda::das_tool=1.1.3" : null) + conda (params.enable_conda ? "bioconda::das_tool=1.1.4" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/das_tool:1.1.3--r41hdfd78af_0' : - 'quay.io/biocontainers/das_tool:1.1.3--r41hdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/das_tool:1.1.4--r41hdfd78af_1' : + 'quay.io/biocontainers/das_tool:1.1.4--r41hdfd78af_1' }" input: tuple val(meta), path(fasta) val(extension) output: - tuple val(meta), path("*.tsv"), emit: scaffolds2bin - path "versions.yml" , emit: versions + tuple val(meta), path("*.tsv"), emit: fastatocontig2bin + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -22,11 +22,12 @@ process DASTOOL_SCAFFOLDS2BIN { def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" def file_extension = extension ? extension : "fasta" - + def clean_fasta = fasta.toString() - ".gz" + def decompress_fasta = fasta.toString() == clean_fasta ? "" : "gunzip -q -f $fasta" """ - gunzip -f *.${file_extension}.gz + $decompress_fasta - Fasta_to_Scaffolds2Bin.sh \\ + Fasta_to_Contig2Bin.sh \\ $args \\ -i . \\ -e $file_extension \\ @@ -34,7 +35,7 @@ process DASTOOL_SCAFFOLDS2BIN { cat <<-END_VERSIONS > versions.yml "${task.process}": - dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool version //' ) + dastool: \$( DAS_Tool --version 2>&1 | grep "DAS Tool" | sed 's/DAS Tool //' ) END_VERSIONS """ } diff --git a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml b/modules/nf-core/modules/dastool/fastatocontig2bin/meta.yml similarity index 83% rename from modules/nf-core/modules/dastool/scaffolds2bin/meta.yml rename to modules/nf-core/modules/dastool/fastatocontig2bin/meta.yml index 0bf8618d..1176ae96 100644 --- a/modules/nf-core/modules/dastool/scaffolds2bin/meta.yml +++ b/modules/nf-core/modules/dastool/fastatocontig2bin/meta.yml @@ -1,4 +1,4 @@ -name: dastool_scaffolds2bin +name: dastool_fastatocontig2bin description: Helper script to convert a set of bins in fasta format to tabular scaffolds2bin format keywords: - binning @@ -31,13 +31,10 @@ input: - fasta: type: file description: Fasta of list of fasta files recommended to be gathered via with .collect() of bins - pattern: "*.{fa,fas,fasta}" - - binner: - type: val - description: Name of the binning software (optional) + pattern: "*.{fa,fa.gz,fas,fas.gz,fna,fna.gz,fasta,fasta.gz}" - extension: type: val - description: Fasta file extension (fa | fas | fasta | ...) + description: Fasta file extension (fa | fas | fasta | ...), without .gz suffix, if gzipped input. output: - meta: @@ -49,10 +46,11 @@ output: type: file description: File containing software versions pattern: "versions.yml" - - scaffolds2bin: + - fastatocontig2bin: type: file - description: tabular scaffolds2bin file for DAS tool input - pattern: "*.scaffolds2bin.tsv" + description: tabular contig2bin file for DAS tool input + pattern: "*.tsv" authors: - "@maxibor" + - "@jfy133" diff --git a/nextflow.config b/nextflow.config index 0583d448..b6646e84 100644 --- a/nextflow.config +++ b/nextflow.config @@ -85,9 +85,10 @@ params { lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // binning options - skip_metabat2 = false - skip_maxbin2 = false - refine_bins_dastool = false + skip_metabat2 = false + skip_maxbin2 = false + refine_bins_dastool = false + refine_bins_dastool_threshold = 0.5 // Bin QC skip_busco = false diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 7c57c7ae..c4c75964 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -52,7 +52,7 @@ workflow BINNING { [ meta_new, depths ] } - ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions) + ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions.first()) // combine depths back with assemblies ch_metabat2_input = assemblies @@ -82,12 +82,12 @@ workflow BINNING { // run binning if ( !params.skip_metabat2 ) { METABAT2_METABAT2 ( ch_metabat2_input ) - ch_versions = ch_versions.mix(METABAT2_METABAT2.out.versions) + ch_versions = ch_versions.mix(METABAT2_METABAT2.out.versions.first()) } if ( !params.skip_maxbin2 ) { MAXBIN2 ( ch_maxbin2_input ) - ch_versions = ch_versions.mix(MAXBIN2.out.versions) + ch_versions = ch_versions.mix(MAXBIN2.out.versions.first()) } // split fastq files, depending @@ -108,7 +108,7 @@ workflow BINNING { ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() - ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) + ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions.first()) // mix all bins together ch_final_bins_for_gunzip = ch_metabat2_results_transposed @@ -116,11 +116,11 @@ workflow BINNING { GUNZIP_BINS ( ch_final_bins_for_gunzip ) ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip - ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions) + ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions.first()) GUNZIP_UNBINS ( ch_split_fasta_results_transposed ) ch_splitfasta_results_gunzipped = GUNZIP_UNBINS.out.gunzip - ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions) + ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions.first()) // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta for grouping to mix back with original depth @@ -136,7 +136,7 @@ workflow BINNING { .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) MAG_DEPTHS ( ch_depth_input ) - ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) + ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions.first()) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples @@ -155,8 +155,8 @@ workflow BINNING { MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) - ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.versions) - ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions) + ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.first()) + ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions.first()) // Group final binned contigs per sample for final output ch_binning_results_gunzipped_final = ch_binning_results_gunzipped diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index f8d7ae9c..59798f61 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -2,8 +2,8 @@ * Binning with MetaBAT2 and MaxBin2 */ -include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' -include { DASTOOL_SCAFFOLDS2BIN as DASTOOL_SCAFFOLDS2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/scaffolds2bin/main.nf' +include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' +include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' workflow BINNING_REFINEMENT { @@ -23,25 +23,42 @@ workflow BINNING_REFINEMENT { .dump(tag: "binrefine_contigs") bins.dump(tag: "binrefine_bins") - ch_bins_for_scaffolds2bin = bins + ch_bins_for_fastatocontig2bin = bins .branch { metabat2: it[0]['binner'] == 'MetaBAT2' maxbin2: it[0]['binner'] == 'MaxBin2' } // Run on each bin file separately - DASTOOL_SCAFFOLDS2BIN_METABAT2 ( ch_bins_for_scaffolds2bin.metabat2, "fa") - DASTOOL_SCAFFOLDS2BIN_MAXBIN2 ( ch_bins_for_scaffolds2bin.maxbin2, "fasta") + DASTOOL_FASTATOCONTIG2BIN_METABAT2 ( ch_bins_for_fastatocontig2bin.metabat2, "fa") + DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 ( ch_bins_for_fastatocontig2bin.maxbin2, "fasta") - ch_versions = ch_versions.mix(DASTOOL_SCAFFOLDS2BIN_METABAT2.out.versions) - ch_versions = ch_versions.mix(DASTOOL_SCAFFOLDS2BIN_MAXBIN2.out.versions) + ch_fastatocontig2bin_for_dastool = Channel.empty() + ch_fastatocontig2bin_for_dastool = ch_fastatocontig2bin_for_dastool + .mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.fastatocontig2bin) + .mix(DASTOOL_FASTATOCONTIG2BIN_MAXBIN2.out.fastatocontig2bin) + .map { + meta, fastatocontig2bin -> + def meta_new = meta.clone() + meta_new.remove('binner') + + [ meta_new, fastatocontig2bin ] + } + .groupTuple(by: 0) + .dump(tag: "fastatocontig2bin_out_mixed") + + ch_input_for_dastool = ch_contigs_for_dastool.join(ch_fastatocontig2bin_for_dastool, by: 0).dump(tag: "input_to_dastool") + + ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.versions.first()) + ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_MAXBIN2.out.versions.first()) // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) // Run DAStool - //DASTOOL_DASTOOL() - //ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions) + DASTOOL_DASTOOL(ch_input_for_dastool, [], []) + ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions.first()) emit: - versions = ch_versions + dastool = DASTOOL_DASTOOL.out.bins + versions = ch_versions } diff --git a/workflows/mag.nf b/workflows/mag.nf index 08d170c5..a267a187 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -248,7 +248,7 @@ workflow MAG { ch_short_reads = Channel.empty() ch_short_reads = ch_short_reads.mix(ADAPTERREMOVAL_SE.out.singles_truncated, ch_adapterremoval_pe_out) - ch_versions = ch_versions.mix(ADAPTERREMOVAL_PE.out.versions, ADAPTERREMOVAL_SE.out.versions) + ch_versions = ch_versions.mix(ADAPTERREMOVAL_PE.out.versions.first(), ADAPTERREMOVAL_SE.out.versions.first()) } From e454bb0f41d40590b0babc040c20df13ba37228e Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Mon, 4 Apr 2022 13:50:22 +0200 Subject: [PATCH 143/246] Update DAS_Tool to allow missing output files --- conf/modules.config | 2 +- conf/test.config | 1 + conf/test_hybrid.config | 2 -- modules.json | 2 +- modules/nf-core/modules/dastool/dastool/main.nf | 4 ++-- 5 files changed, 5 insertions(+), 6 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index baeb97f4..b901b098 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -476,8 +476,8 @@ process { ], ] ext.prefix = { "${meta.assembler}-${meta.id}" } - // TODO move penalties to test config if they work! ext.args = "--write_bins --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" + } withName: CUSTOM_DUMPSOFTWAREVERSIONS { diff --git a/conf/test.config b/conf/test.config index 5df32bdb..9ca02d83 100644 --- a/conf/test.config +++ b/conf/test.config @@ -28,4 +28,5 @@ params { max_unbinned_contigs = 2 busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" gtdb = false + refine_bins_dastool = true } diff --git a/conf/test_hybrid.config b/conf/test_hybrid.config index 6ba74bfb..edc879e7 100644 --- a/conf/test_hybrid.config +++ b/conf/test_hybrid.config @@ -25,6 +25,4 @@ params { max_unbinned_contigs = 2 busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" gtdb = false - refine_bins_dastool = true - refine_bins_dastool_threshold = 0 } diff --git a/modules.json b/modules.json index 69f7927f..c37ac3a1 100644 --- a/modules.json +++ b/modules.json @@ -19,7 +19,7 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "dastool/dastool": { - "git_sha": "8ce68107871c96519b3eb0095d97896e34ef4489" + "git_sha": "ae48653bd2d169510580220bb62d96f830c31293" }, "dastool/fastatocontig2bin": { "git_sha": "8ce68107871c96519b3eb0095d97896e34ef4489" diff --git a/modules/nf-core/modules/dastool/dastool/main.nf b/modules/nf-core/modules/dastool/dastool/main.nf index 968f85de..a7d9c6f6 100644 --- a/modules/nf-core/modules/dastool/dastool/main.nf +++ b/modules/nf-core/modules/dastool/dastool/main.nf @@ -14,8 +14,8 @@ process DASTOOL_DASTOOL { output: tuple val(meta), path("*.log") , emit: log - tuple val(meta), path("*_summary.tsv") , emit: summary - tuple val(meta), path("*_DASTool_contig2bin.tsv") , emit: contig2bin + tuple val(meta), path("*_summary.tsv") , optional: true, emit: summary + tuple val(meta), path("*_DASTool_contig2bin.tsv") , optional: true, emit: contig2bin tuple val(meta), path("*.eval") , optional: true, emit: eval tuple val(meta), path("*_DASTool_bins/*.fa") , optional: true, emit: bins tuple val(meta), path("*.pdf") , optional: true, emit: pdfs From 5477e45e571713b4029d62be2a48ee180f8e24a1 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 7 Apr 2022 13:47:44 +0200 Subject: [PATCH 144/246] Finalise DAS_Tool - fixing version reporting and add documentation --- CHANGELOG.md | 1 + conf/modules.config | 26 ++++++++------------------ conf/test.config | 1 + docs/output.md | 17 +++++++++++++++++ docs/usage.md | 12 ++++++++++++ modules/local/convert_depths.nf | 1 + modules/local/split_fasta.nf | 2 +- nextflow_schema.json | 31 +++++++++++++++++++++++++++---- subworkflows/local/binning.nf | 11 ++++++----- workflows/mag.nf | 10 +++++----- 10 files changed, 79 insertions(+), 33 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index be27e2d5..6cdc5410 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,6 +11,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool - [#284](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer +- [#284](https://github.com/nf-core/mag/pull/291) - Add DAS_Tool for bin refinement ### `Changed` diff --git a/conf/modules.config b/conf/modules.config index b901b098..d5f90824 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -444,27 +444,11 @@ process { } withName: DASTOOL_FASTATOCONTIG2BIN_METABAT2 { - publishDir = [ - [ - path: { "${params.outdir}/BinRefinement/fastatocontig2bin" }, - mode: params.publish_dir_mode, - pattern: '*.tsv' - ], - ] ext.prefix = { "${meta.assembler}-MetaBAT2-${meta.id}" } - } withName: DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 { - publishDir = [ - [ - path: { "${params.outdir}/BinRefinement/fastatocontig2bin" }, - mode: params.publish_dir_mode, - pattern: '*.tsv' - ], - ] ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } - } withName: DASTOOL_DASTOOL { @@ -472,12 +456,18 @@ process { [ path: { "${params.outdir}/BinRefinement/" }, mode: params.publish_dir_mode, - pattern: '*.tsv' + pattern: '*.{tsv,log,eval,seqlength}' + ], + [ + path: { "${params.outdir}/BinRefinement/" }, + mode: params.publish_dir_mode, + pattern: '*_bins/*.fa' ], ] ext.prefix = { "${meta.assembler}-${meta.id}" } ext.args = "--write_bins --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" - + // if SCGs not found, bins cannot be assigned and DAS_tool will die + errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : task.exitStatus == 1 ? 'ignore' : 'finish' } } withName: CUSTOM_DUMPSOFTWAREVERSIONS { diff --git a/conf/test.config b/conf/test.config index 9ca02d83..52e4af98 100644 --- a/conf/test.config +++ b/conf/test.config @@ -29,4 +29,5 @@ params { busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" gtdb = false refine_bins_dastool = true + refine_bins_dastool_threshold = 0 } diff --git a/docs/output.md b/docs/output.md index 48b2abd5..ff17a364 100644 --- a/docs/output.md +++ b/docs/output.md @@ -404,6 +404,23 @@ Besides the reference files or output files created by BUSCO, the following summ
+### Bin Refinement + +[DAS Tool](https://github.com/cmks/DAS_Tool) is an automated method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. MAG uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins. + +
+Output files + +- `BinRefinement/` + + - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets + - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets + - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners + - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information + - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig + +
+ ## Taxonomic classification of binned genomes ### CAT diff --git a/docs/usage.md b/docs/usage.md index ac4bab6e..0c6a4e72 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -287,3 +287,15 @@ NXF_OPTS='-Xms1g -Xmx4g' MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: [Characteristic patterns of ancient DNA (aDNA) damage](<(https://doi.org/10.1073/pnas.0704665104)>), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. + +## A note on bin refinement + +DAS_Tool may not always be able to refine bins due to insufficient recovery of enough single-copy genes. In these cases you will get a NOTE such as + +```console +[16/d330a6] NOTE: Process `NFCORE_MAG:MAG:BINNING_REFINEMENT:DASTOOL_DASTOOL (test_minigut_sample2)` terminated with an error exit status (1) -- Error is ignored +``` + +In this case, DAS_Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the BinRefinement results directory for that sample. + +If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS_Tool publication](https://www.nature.com/articles/s41564-018-0171-1). diff --git a/modules/local/convert_depths.nf b/modules/local/convert_depths.nf index dcebe79a..a307f42f 100644 --- a/modules/local/convert_depths.nf +++ b/modules/local/convert_depths.nf @@ -12,6 +12,7 @@ process CONVERT_DEPTHS { output: // need to add empty val because representing reads as we dont want maxbin to calculate for us. tuple val(meta), path(fasta), val([]), path("*_mb2_depth.txt"), emit: output + path "versions.yml" , emit: versions script: def prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/local/split_fasta.nf b/modules/local/split_fasta.nf index 2d27311e..15e0a5e7 100644 --- a/modules/local/split_fasta.nf +++ b/modules/local/split_fasta.nf @@ -15,7 +15,7 @@ process SPLIT_FASTA { tuple val(meta), path("*.[0-9]*.fa.gz") , optional:true, emit: unbinned tuple val(meta), path("*.pooled.fa.gz") , optional:true, emit: pooled tuple val(meta), path("*.remaining.fa.gz") , optional:true, emit: remaining - path "versions.yml" , emit: versions + path "versions.yml" , emit: versions script: """ diff --git a/nextflow_schema.json b/nextflow_schema.json index b651e95a..2c0ec394 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,10 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": ["input", "outdir"], + "required": [ + "input", + "outdir" + ], "properties": { "input": { "type": "string", @@ -170,7 +173,14 @@ "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", - "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], + "enum": [ + "symlink", + "rellink", + "link", + "copy", + "copyNoFollow", + "move" + ], "hidden": true }, "email_on_fail": { @@ -278,7 +288,10 @@ "type": "string", "default": "fastp", "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", - "enum": ["fastp", "adapterremoval"] + "enum": [ + "fastp", + "adapterremoval" + ] }, "reads_minlength": { "type": "integer", @@ -605,6 +618,16 @@ "type": "boolean", "description": "Save the used BUSCO lineage datasets provided via --busco_reference or downloaded when not using --busco_reference or --busco_download_path.", "help_text": "Useful to allow reproducibility, as BUSCO datasets are frequently updated and old versions do not always remain accessible." + }, + "refine_bins_dastool": { + "type": "boolean", + "description": "Turn on bin refinement using DAS_Tool." + }, + "refine_bins_dastool_threshold": { + "type": "number", + "default": 0.5, + "description": "Specify single-copy gene score threshold for bin refinement.", + "help_text": "Score threshold for single-copy gene selection algorithm to keep selecting bins, with a value ranging from 0-1.\n\nFor description of scoring algorithm, see: Sieber, Christian M. K., et al. 2018. Nature Microbiology 3 (7): 836\u201343. https://doi.org/10.1038/s41564-018-0171-1.\n\n> Modifies DAS_Tool parameter --score_threshold\n" } } }, @@ -700,4 +723,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} +} \ No newline at end of file diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index c4c75964..10d65dd2 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -77,6 +77,7 @@ workflow BINNING { [ meta_new, assembly, reads, depth ] } + ch_versions = ch_versions.mix(CONVERT_DEPTHS.out.versions.first()) } // run binning @@ -108,7 +109,7 @@ workflow BINNING { ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() - ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions.first()) + ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) // mix all bins together ch_final_bins_for_gunzip = ch_metabat2_results_transposed @@ -136,7 +137,7 @@ workflow BINNING { .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) MAG_DEPTHS ( ch_depth_input ) - ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions.first()) + ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) // create file containing group information for all samples @@ -155,8 +156,8 @@ workflow BINNING { MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY ( MAG_DEPTHS.out.depths.map{it[1]}.collect() ) - ch_versions = ch_versions.mix(MAG_DEPTHS_PLOT.out.first()) - ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions.first()) + ch_versions = ch_versions.mix( MAG_DEPTHS_PLOT.out.versions ) + ch_versions = ch_versions.mix( MAG_DEPTHS_SUMMARY.out.versions ) // Group final binned contigs per sample for final output ch_binning_results_gunzipped_final = ch_binning_results_gunzipped @@ -173,5 +174,5 @@ workflow BINNING { unbinned = ch_splitfasta_results_gunzipped.groupTuple() unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary - versions = ch_versions + versions = ch_versions.dump(tag: "binning_versions") } diff --git a/workflows/mag.nf b/workflows/mag.nf index a267a187..418d751a 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -649,6 +649,7 @@ workflow MAG { */ if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) + ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) } } @@ -683,13 +684,12 @@ workflow MAG { ) */ - //This is the local module because (1) fastq file name clashes [probably solveable] and (2) host removal bowtie reporting - ch_multiqc_additional = Channel.empty() + ch_multiqc_readprep = Channel.empty() if ( params.clip_tool == "fastp") { - ch_multiqc_additional = ch_multiqc_additional.mix(FASTP.out.json.collect{it[1]}.ifEmpty([])) + ch_multiqc_readprep = ch_multiqc_readprep.mix(FASTP.out.json.collect{it[1]}.ifEmpty([])) } else if ( params.clip_tool == "adapterremoval" ) { - ch_multiqc_additional = ch_multiqc_additional.mix(ADAPTERREMOVAL_PE.out.log.collect{it[1]}.ifEmpty([]), ADAPTERREMOVAL_SE.out.log.collect{it[1]}.ifEmpty([])) + ch_multiqc_readprep = ch_multiqc_readprep.mix(ADAPTERREMOVAL_PE.out.log.collect{it[1]}.ifEmpty([]), ADAPTERREMOVAL_SE.out.log.collect{it[1]}.ifEmpty([])) } MULTIQC ( @@ -701,7 +701,7 @@ workflow MAG { ch_quast_multiqc.collect().ifEmpty([]), ch_bowtie2_assembly_multiqc.collect().ifEmpty([]), ch_busco_multiqc.collect().ifEmpty([]), - ch_multiqc_additional.collect().ifEmpty([]), + ch_multiqc_readprep.collect().ifEmpty([]), ) multiqc_report = MULTIQC.out.report.toList() ch_versions = ch_versions.mix(MULTIQC.out.versions) From dc2dd14cd5f77137dba1375c299583781d6fe8c6 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 7 Apr 2022 13:49:30 +0200 Subject: [PATCH 145/246] Add missing output directory to OUTPUT docs --- docs/output.md | 1 + 1 file changed, 1 insertion(+) diff --git a/docs/output.md b/docs/output.md index ff17a364..a45873cd 100644 --- a/docs/output.md +++ b/docs/output.md @@ -418,6 +418,7 @@ Besides the reference files or output files created by BUSCO, the following summ - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig + - `[assembler]-[sample/group]_DASTool_bins/*`: Directory containing bins in FASTA format From dc26f8d2a23c1f6839876c14c07da05a6b7167cf Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 7 Apr 2022 13:51:36 +0200 Subject: [PATCH 146/246] prettier --- modules.json | 2 +- nextflow_schema.json | 21 ++++----------------- 2 files changed, 5 insertions(+), 18 deletions(-) diff --git a/modules.json b/modules.json index c37ac3a1..f09b78c5 100644 --- a/modules.json +++ b/modules.json @@ -62,4 +62,4 @@ } } } -} \ No newline at end of file +} diff --git a/nextflow_schema.json b/nextflow_schema.json index 2c0ec394..3b6d97d5 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,10 +10,7 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": [ - "input", - "outdir" - ], + "required": ["input", "outdir"], "properties": { "input": { "type": "string", @@ -173,14 +170,7 @@ "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", - "enum": [ - "symlink", - "rellink", - "link", - "copy", - "copyNoFollow", - "move" - ], + "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], "hidden": true }, "email_on_fail": { @@ -288,10 +278,7 @@ "type": "string", "default": "fastp", "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", - "enum": [ - "fastp", - "adapterremoval" - ] + "enum": ["fastp", "adapterremoval"] }, "reads_minlength": { "type": "integer", @@ -723,4 +710,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} \ No newline at end of file +} From c8bb385ef8a77f9cd4ee93fd81560b829fe27745 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 7 Apr 2022 14:45:13 +0200 Subject: [PATCH 147/246] Update README --- README.md | 1 + 1 file changed, 1 insertion(+) diff --git a/README.md b/README.md index 59fe2224..7f69d209 100644 --- a/README.md +++ b/README.md @@ -37,6 +37,7 @@ The pipeline then: - (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) - predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) - performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/) and/or with [MaxBin2](https://sourceforge.net/projects/maxbin2/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) +- optionally refines bins with [DAS_Tool](https://github.com/cmks/DAS_Tool) - assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. From 78b1ed34c7c41c2018df408a4f8d0e79ec05ceee Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Fri, 8 Apr 2022 09:22:58 +0200 Subject: [PATCH 148/246] Apply suggestions from code review Co-authored-by: Sabrina Krakau Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- docs/output.md | 3 +-- subworkflows/local/binning.nf | 2 +- subworkflows/local/binning_refinement.nf | 4 ---- workflows/mag.nf | 2 -- 4 files changed, 2 insertions(+), 9 deletions(-) diff --git a/docs/output.md b/docs/output.md index a45873cd..e82ace68 100644 --- a/docs/output.md +++ b/docs/output.md @@ -406,13 +406,12 @@ Besides the reference files or output files created by BUSCO, the following summ ### Bin Refinement -[DAS Tool](https://github.com/cmks/DAS_Tool) is an automated method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. MAG uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins. +[DAS Tool](https://github.com/cmks/DAS_Tool) is an automated method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. nf-core/mag uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins.
Output files - `BinRefinement/` - - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 10d65dd2..0b5b0c9c 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -174,5 +174,5 @@ workflow BINNING { unbinned = ch_splitfasta_results_gunzipped.groupTuple() unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary - versions = ch_versions.dump(tag: "binning_versions") + versions = ch_versions } diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 59798f61..4fa12cdb 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -20,8 +20,6 @@ workflow BINNING_REFINEMENT { def meta_new = meta.clone() [ meta_new, assembly ] } - .dump(tag: "binrefine_contigs") - bins.dump(tag: "binrefine_bins") ch_bins_for_fastatocontig2bin = bins .branch { @@ -41,11 +39,9 @@ workflow BINNING_REFINEMENT { meta, fastatocontig2bin -> def meta_new = meta.clone() meta_new.remove('binner') - [ meta_new, fastatocontig2bin ] } .groupTuple(by: 0) - .dump(tag: "fastatocontig2bin_out_mixed") ch_input_for_dastool = ch_contigs_for_dastool.join(ch_fastatocontig2bin_for_dastool, by: 0).dump(tag: "input_to_dastool") diff --git a/workflows/mag.nf b/workflows/mag.nf index 418d751a..4d8822dc 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -654,8 +654,6 @@ workflow MAG { } - - CUSTOM_DUMPSOFTWAREVERSIONS ( ch_versions.unique().collectFile(name: 'collated_versions.yml') ) From e7266eba32604883cc5c3348ff3634dc95ad0998 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 8 Apr 2022 14:50:48 +0200 Subject: [PATCH 149/246] Attempt at allowing user to define which binning files go into downstream --- conf/base.config | 10 ++++++ conf/modules.config | 5 +-- docs/output.md | 2 +- docs/usage.md | 2 ++ modules/local/rename_dastool.nf | 28 +++++++++++++++++ nextflow.config | 3 +- nextflow_schema.json | 32 ++++++++++++++++--- subworkflows/local/binning_refinement.nf | 22 ++++++++++--- workflows/mag.nf | 39 +++++++++++++++--------- 9 files changed, 114 insertions(+), 29 deletions(-) create mode 100644 modules/local/rename_dastool.nf diff --git a/conf/base.config b/conf/base.config index 38dc0ad7..2abc49b5 100644 --- a/conf/base.config +++ b/conf/base.config @@ -159,4 +159,14 @@ process { cpus = { check_max (8 * task.attempt, 'cpus' ) } memory = { check_max (20.GB * task.attempt, 'memory' ) } } + + withName: MAXBIN2 { + // often fails when insufficient information, so we allow it to gracefully fail without filling the pipeline + errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } + } + + withName: DASTOOL_DASTOOL { + // if SCGs not found, bins cannot be assigned and DAS_tool will die + errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : task.exitStatus == 1 ? 'ignore' : 'finish' } + } } diff --git a/conf/modules.config b/conf/modules.config index d5f90824..e12e1bec 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -420,7 +420,6 @@ process { ext.prefix = { "${meta.assembler}-MaxBin2-${meta.id}" } // if no gene found, will crash so allow ignore so rest of pipeline // completes but without MaxBin2 results - errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } } withName: SPLIT_FASTA { @@ -465,9 +464,7 @@ process { ], ] ext.prefix = { "${meta.assembler}-${meta.id}" } - ext.args = "--write_bins --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" - // if SCGs not found, bins cannot be assigned and DAS_tool will die - errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : task.exitStatus == 1 ? 'ignore' : 'finish' } + ext.args = "--write_bins --write_unbinned --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" } withName: CUSTOM_DUMPSOFTWAREVERSIONS { diff --git a/docs/output.md b/docs/output.md index e82ace68..472b1d2a 100644 --- a/docs/output.md +++ b/docs/output.md @@ -425,7 +425,7 @@ Besides the reference files or output files created by BUSCO, the following summ ### CAT -[CAT](https://github.com/dutilh/CAT) is a toolkit for annotating contigs and bins from metagenome-assembled-genomes. The MAG pipeline uses CAT to assign taxonomy to genome bins based on the taxnomy of the contigs. +[CAT](https://github.com/dutilh/CAT) is a toolkit for annotating contigs and bins from metagenome-assembled-genomes. The nf-core/mag pipeline uses CAT to assign taxonomy to genome bins based on the taxnomy of the contigs.
Output files diff --git a/docs/usage.md b/docs/usage.md index 0c6a4e72..1198aaa0 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -299,3 +299,5 @@ DAS_Tool may not always be able to refine bins due to insufficient recovery of e In this case, DAS_Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the BinRefinement results directory for that sample. If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS_Tool publication](https://www.nature.com/articles/s41564-018-0171-1). + +Furthermore, only diff --git a/modules/local/rename_dastool.nf b/modules/local/rename_dastool.nf new file mode 100644 index 00000000..c846c67d --- /dev/null +++ b/modules/local/rename_dastool.nf @@ -0,0 +1,28 @@ +process RENAME_DASTOOL { + tag "${meta.assembler}-${meta.id}" + label 'process_low' + + // Using container from multiqc since it'll be included anyway + conda (params.enable_conda ? "bioconda::multiqc=1.12" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.12--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.12--pyhdfd78af_0' }" + + input: + tuple val(meta), path(bins) + + output: + tuple val(meta), path("*_refined.fasta") , optional:true, emit: refined_bins + tuple val(meta), path("*_refined_unbinned.fasta") , optional:true, emit: refined_unbins + + script: + """ + if [[ -f unbinned.fa ]]; then + mv unbinned.fa ${meta.assembler}-${meta.id}_refined_unbinned.fasta + fi + + for i in *.fa; do + mv \$i \${i%%.fa}_refined.fasta + done + """ +} diff --git a/nextflow.config b/nextflow.config index b6646e84..e7d7f11c 100644 --- a/nextflow.config +++ b/nextflow.config @@ -81,7 +81,7 @@ params { longreads_min_length = 1000 longreads_keep_percent = 90 longreads_length_weight = 10 - // lambda_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Escherichia_virus_Lambda/all_assembly_versions/GCA_000840245.1_ViralProj14204/GCA_000840245.1_ViralProj14204_genomic.fna.gz" + // lambda_reference = "ftp://ftp.ncbi.nlm.nih.gov/genomes/genbank/viral/Escherichia_virus_Lambda/all_assembly_versions/GCA_000840245.1_ViralProj14204/GCA_000840245.1_ViralProj14204_genomic.fna.gz" lambda_reference = "${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz" // binning options @@ -89,6 +89,7 @@ params { skip_maxbin2 = false refine_bins_dastool = false refine_bins_dastool_threshold = 0.5 + postbinning_input = 'raw_bins_only' // Bin QC skip_busco = false diff --git a/nextflow_schema.json b/nextflow_schema.json index 3b6d97d5..a71f9f92 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,10 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": ["input", "outdir"], + "required": [ + "input", + "outdir" + ], "properties": { "input": { "type": "string", @@ -170,7 +173,14 @@ "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", - "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], + "enum": [ + "symlink", + "rellink", + "link", + "copy", + "copyNoFollow", + "move" + ], "hidden": true }, "email_on_fail": { @@ -278,7 +288,10 @@ "type": "string", "default": "fastp", "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", - "enum": ["fastp", "adapterremoval"] + "enum": [ + "fastp", + "adapterremoval" + ] }, "reads_minlength": { "type": "integer", @@ -615,6 +628,17 @@ "default": 0.5, "description": "Specify single-copy gene score threshold for bin refinement.", "help_text": "Score threshold for single-copy gene selection algorithm to keep selecting bins, with a value ranging from 0-1.\n\nFor description of scoring algorithm, see: Sieber, Christian M. K., et al. 2018. Nature Microbiology 3 (7): 836\u201343. https://doi.org/10.1038/s41564-018-0171-1.\n\n> Modifies DAS_Tool parameter --score_threshold\n" + }, + "postbinning_input": { + "type": "string", + "default": "raw_bins_only", + "description": "Specify which binning output is sent for downstream annotation, taxonomic categorisation, assembly qualiy control etc.", + "help_text": "`raw_bins_only`: only bins (and unbinned contigs) from the binners.\n`refined_bins_only`: only bins (and unbinned contigs) from the bin refinement step .\n`both`: bins and and unbinned contigs from both the binning and bin refinement steps.", + "enum": [ + "raw_bins_only", + "refined_bins_only", + "both" + ] } } }, @@ -710,4 +734,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} +} \ No newline at end of file diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 4fa12cdb..7774a02e 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -4,7 +4,9 @@ include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' -include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' +include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' +include { RENAME_DASTOOL } from '../../modules/local/rename_dastool' + workflow BINNING_REFINEMENT { take: @@ -48,13 +50,25 @@ workflow BINNING_REFINEMENT { ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.versions.first()) ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_MAXBIN2.out.versions.first()) - // Concatenate each binner table per sample (based on contig names in FASTA file, not fasta file itself, so should be OK) - // Run DAStool DASTOOL_DASTOOL(ch_input_for_dastool, [], []) ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions.first()) + ch_input_for_renamedastool = DASTOOL_DASTOOL.out.bins + .map { + meta, bins -> + def meta_new = meta.clone() + meta_new['binner'] = 'DASTool' + [ meta_new, bins ] + } + + RENAME_DASTOOL ( ch_input_for_renamedastool ) + + RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool_bins") + RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool_unbins") + emit: - dastool = DASTOOL_DASTOOL.out.bins + refined_bins = RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool") + refined_unbins = RENAME_DASTOOL.out.refined_unbins versions = ch_versions } diff --git a/workflows/mag.nf b/workflows/mag.nf index 4d8822dc..917b8d06 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -549,11 +549,30 @@ workflow MAG { ch_versions = ch_versions.mix(BINNING_PREPARATION.out.bowtie2_version.first()) ch_versions = ch_versions.mix(BINNING.out.versions) + /* + * DAS_Tool: binning refinement + */ + if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { + BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) + ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) + } + + if ( params.postbinning_input == 'raw_bins_only' ) { + ch_input_for_postbinning_bins = BINNING.out..dump(tag: "bin_refine_out_raw") + ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) + } else if ( params.postbinning_input == 'refined_bins_only' ) { + ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins.dump(tag: "bin_refine_out_refinedonly") + ch_input_for_postbinning_bins_unbins = BINNING_REFINEMENT.out.refined_bins.mix(BINNING_REFINEMENT.out.refined_unbins) + } else if (params.postbinning_input == 'both') { + ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins).dump(tag: "bin_refine_out_both") + ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.bins,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) + } + if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly */ - ch_input_bins_busco = BINNING.out.bins.mix( BINNING.out.unbinned ).transpose() + ch_input_bins_busco = ch_input_for_postbinning_bins_unbins.transpose() BUSCO_QC ( ch_busco_db_file, ch_busco_download_folder, @@ -571,8 +590,7 @@ workflow MAG { ch_quast_bins_summary = Channel.empty() if (!params.skip_quast){ - ch_input_for_quast_bins = BINNING.out.bins - .mix( BINNING.out.unbinned ) + ch_input_for_quast_bins = ch_input_for_postbinning_bins_unbins .groupTuple() .map{ meta, reads -> @@ -597,7 +615,7 @@ workflow MAG { ch_cat_db = CAT_DB_GENERATE.out.db } CAT ( - BINNING.out.bins, + ch_input_for_postbinning_bins, ch_cat_db ) ch_versions = ch_versions.mix(CAT.out.versions.first()) @@ -608,7 +626,7 @@ workflow MAG { ch_gtdbtk_summary = Channel.empty() if ( gtdb ){ GTDBTK ( - BINNING.out.bins, + ch_input_for_postbinning_bins_unbins, ch_busco_summary, ch_gtdb ) @@ -628,7 +646,7 @@ workflow MAG { /* * Prokka: Genome annotation */ - ch_bins_for_prokka = BINNING.out.bins.mix(BINNING.out.unbinned).transpose() + ch_bins_for_prokka = ch_input_for_postbinning_bins_unbins.transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() @@ -643,15 +661,6 @@ workflow MAG { ) ch_versions = ch_versions.mix(PROKKA.out.versions.first()) } - - /* - * DAS_Tool: binning refinement - */ - if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { - BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) - ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) - } - } CUSTOM_DUMPSOFTWAREVERSIONS ( From 31ddfca88d08733000076e8eba3caf479a61dbd3 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 8 Apr 2022 21:56:14 +0200 Subject: [PATCH 150/246] Adds improved output for bin_refinement to include unbins --- subworkflows/local/binning_refinement.nf | 5 +---- workflows/mag.nf | 6 +++--- 2 files changed, 4 insertions(+), 7 deletions(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 7774a02e..c6074bdf 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -64,11 +64,8 @@ workflow BINNING_REFINEMENT { RENAME_DASTOOL ( ch_input_for_renamedastool ) - RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool_bins") - RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool_unbins") - emit: - refined_bins = RENAME_DASTOOL.out.refined_bins.dump(tag: "out_from_renamedastool") + refined_bins = RENAME_DASTOOL.out.refined_bins refined_unbins = RENAME_DASTOOL.out.refined_unbins versions = ch_versions } diff --git a/workflows/mag.nf b/workflows/mag.nf index 917b8d06..604c4de2 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -558,13 +558,13 @@ workflow MAG { } if ( params.postbinning_input == 'raw_bins_only' ) { - ch_input_for_postbinning_bins = BINNING.out..dump(tag: "bin_refine_out_raw") + ch_input_for_postbinning_bins = BINNING.out.bins ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) } else if ( params.postbinning_input == 'refined_bins_only' ) { - ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins.dump(tag: "bin_refine_out_refinedonly") + ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins ch_input_for_postbinning_bins_unbins = BINNING_REFINEMENT.out.refined_bins.mix(BINNING_REFINEMENT.out.refined_unbins) } else if (params.postbinning_input == 'both') { - ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins).dump(tag: "bin_refine_out_both") + ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.bins,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) } From fc66b5c7ca86955f3335b943aca7987f0b26a1e8 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 8 Apr 2022 22:00:35 +0200 Subject: [PATCH 151/246] Prettier --- nextflow_schema.json | 27 +++++---------------------- 1 file changed, 5 insertions(+), 22 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index a71f9f92..ed5f9bff 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,10 +10,7 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": [ - "input", - "outdir" - ], + "required": ["input", "outdir"], "properties": { "input": { "type": "string", @@ -173,14 +170,7 @@ "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", - "enum": [ - "symlink", - "rellink", - "link", - "copy", - "copyNoFollow", - "move" - ], + "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], "hidden": true }, "email_on_fail": { @@ -288,10 +278,7 @@ "type": "string", "default": "fastp", "description": "Specify which adapter clipping tool to use. Options: 'fastp', 'adapterremoval'", - "enum": [ - "fastp", - "adapterremoval" - ] + "enum": ["fastp", "adapterremoval"] }, "reads_minlength": { "type": "integer", @@ -634,11 +621,7 @@ "default": "raw_bins_only", "description": "Specify which binning output is sent for downstream annotation, taxonomic categorisation, assembly qualiy control etc.", "help_text": "`raw_bins_only`: only bins (and unbinned contigs) from the binners.\n`refined_bins_only`: only bins (and unbinned contigs) from the bin refinement step .\n`both`: bins and and unbinned contigs from both the binning and bin refinement steps.", - "enum": [ - "raw_bins_only", - "refined_bins_only", - "both" - ] + "enum": ["raw_bins_only", "refined_bins_only", "both"] } } }, @@ -734,4 +717,4 @@ "$ref": "#/definitions/ancient_dna_assembly" } ] -} \ No newline at end of file +} From fe14e5331e5a37356709b07e515a1ba9ea5902d5 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 11 Apr 2022 13:54:26 +0200 Subject: [PATCH 152/246] Separate bin refinement CI test --- .github/workflows/ci.yml | 10 +++++++++- conf/test.config | 2 -- conf/test_binrefinement.config | 34 ++++++++++++++++++++++++++++++++++ nextflow.config | 1 + 4 files changed, 44 insertions(+), 3 deletions(-) create mode 100644 conf/test_binrefinement.config diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index dfa368a4..c3688132 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -55,7 +55,15 @@ jobs: matrix: # Run remaining test profiles with minimum nextflow version profile: - [test_host_rm, test_hybrid, test_hybrid_host_rm, test_busco_auto, test_ancient_dna, test_adapterremoval] + [ + test_host_rm, + test_hybrid, + test_hybrid_host_rm, + test_busco_auto, + test_ancient_dna, + test_adapterremoval, + test_binrefinement, + ] steps: - name: Check out pipeline code uses: actions/checkout@v2 diff --git a/conf/test.config b/conf/test.config index 52e4af98..5df32bdb 100644 --- a/conf/test.config +++ b/conf/test.config @@ -28,6 +28,4 @@ params { max_unbinned_contigs = 2 busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" gtdb = false - refine_bins_dastool = true - refine_bins_dastool_threshold = 0 } diff --git a/conf/test_binrefinement.config b/conf/test_binrefinement.config new file mode 100644 index 00000000..1931dd11 --- /dev/null +++ b/conf/test_binrefinement.config @@ -0,0 +1,34 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for running minimal tests +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run nf-core/mag -profile test, --outdir + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Test profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + + // Limit resources so that this can run on GitHub Actions + max_cpus = 2 + max_memory = '6.GB' + max_time = '6.h' + + // Input data + input = 'https://raw.githubusercontent.com/nf-core/test-datasets/mag/samplesheets/samplesheet.csv' + centrifuge_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_cf.tar.gz" + kraken2_db = "https://raw.githubusercontent.com/nf-core/test-datasets/mag/test_data/minigut_kraken.tgz" + skip_krona = true + min_length_unbinned_contigs = 1 + max_unbinned_contigs = 2 + busco_reference = "https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz" + gtdb = false + refine_bins_dastool = true + refine_bins_dastool_threshold = 0 + postbinning_input = 'both' +} diff --git a/nextflow.config b/nextflow.config index e7d7f11c..7f01d694 100644 --- a/nextflow.config +++ b/nextflow.config @@ -217,6 +217,7 @@ profiles { test_full { includeConfig 'conf/test_full.config' } test_ancient_dna { includeConfig 'conf/test_ancient_dna.config' } test_adapterremoval { includeConfig 'conf/test_adapterremoval.config' } + test_binrefinement { includeConfig 'conf/test_binrefinement.config' } } From 532a0b2673ad29b531399a9ba63c7818cada13dd Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 12 Apr 2022 10:40:18 +0200 Subject: [PATCH 153/246] Fix non-binrefinement channel output and add error message --- workflows/mag.nf | 27 +++++++++++++++++---------- 1 file changed, 17 insertions(+), 10 deletions(-) diff --git a/workflows/mag.nf b/workflows/mag.nf index 604c4de2..db6df316 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -35,6 +35,8 @@ for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true // Check mandatory parameters if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input samplesheet not specified!' } +if ( !params.refine_bins_dastool && params.postbinning_input != 'raw_bins_only' ) { exit 1, "[nf-core/mag] error: Can only specify '--postbinning_input ${params.postbinning_input}' for downstream steps if bin refinement activated with --refine_bins_dastool! Check input." } + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ CONFIG FILES @@ -555,19 +557,24 @@ workflow MAG { if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) - } - if ( params.postbinning_input == 'raw_bins_only' ) { - ch_input_for_postbinning_bins = BINNING.out.bins - ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) - } else if ( params.postbinning_input == 'refined_bins_only' ) { - ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins - ch_input_for_postbinning_bins_unbins = BINNING_REFINEMENT.out.refined_bins.mix(BINNING_REFINEMENT.out.refined_unbins) - } else if (params.postbinning_input == 'both') { - ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) - ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.bins,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) + if ( params.postbinning_input == 'raw_bins_only' ) { + ch_input_for_postbinning_bins = BINNING.out.bins + ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) + } else if ( params.postbinning_input == 'refined_bins_only' ) { + ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins + ch_input_for_postbinning_bins_unbins = BINNING_REFINEMENT.out.refined_bins.mix(BINNING_REFINEMENT.out.refined_unbins) + } else if (params.postbinning_input == 'both') { + ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) + ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) + } + + } else { + ch_input_for_postbinning_bins = BINNING.out.bins + ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) } + if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly From e14c3f0b0c4598282ae2e0ad1c4a9fa44242fdee Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 14 Apr 2022 12:51:26 +0200 Subject: [PATCH 154/246] Fix skip-metabat causing undefined out channel and add DASTool as binner to file names via prefix as per @skrakau --- conf/modules.config | 7 ++++++- modules/local/rename_dastool.nf | 2 +- subworkflows/local/binning.nf | 9 +++++++-- 3 files changed, 14 insertions(+), 4 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index e12e1bec..937f80a4 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -463,10 +463,15 @@ process { pattern: '*_bins/*.fa' ], ] - ext.prefix = { "${meta.assembler}-${meta.id}" } + ext.prefix = { "${meta.assembler}-DASTool-${meta.id}" } ext.args = "--write_bins --write_unbinned --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" } + withName: RENAME_DASTOOL { + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + } + + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/modules/local/rename_dastool.nf b/modules/local/rename_dastool.nf index c846c67d..3f7bf6cb 100644 --- a/modules/local/rename_dastool.nf +++ b/modules/local/rename_dastool.nf @@ -18,7 +18,7 @@ process RENAME_DASTOOL { script: """ if [[ -f unbinned.fa ]]; then - mv unbinned.fa ${meta.assembler}-${meta.id}_refined_unbinned.fasta + mv unbinned.fa ${meta.assembler}-${meta.binner}-${meta.id}_refined_unbinned.fasta fi for i in *.fa; do diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 0b5b0c9c..9805d450 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -105,8 +105,13 @@ workflow BINNING { // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for // MAG_DEPTHS, first have to separate and re-group due to limitation of // GUNZIP module - ch_metabat2_results_transposed = METABAT2_METABAT2.out.fasta.transpose() - ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() + if ( !params.skip_metabat2 ) { + ch_metabat2_results_transposed = METABAT2_METABAT2.out.fasta.transpose() + } + + if ( !params.skip_maxbin2 ) { + ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() + } ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) From 2c081df9d80ece27b3fbe6876a7cdd4219ddbef4 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 14 Apr 2022 11:06:38 +0200 Subject: [PATCH 155/246] Add error in case of invalid bin refinement parameter combination --- lib/WorkflowMag.groovy | 11 +++++++++++ 1 file changed, 11 insertions(+) diff --git a/lib/WorkflowMag.groovy b/lib/WorkflowMag.groovy index ce2cf932..33f4ab7b 100755 --- a/lib/WorkflowMag.groovy +++ b/lib/WorkflowMag.groovy @@ -79,6 +79,17 @@ class WorkflowMag { } } + // Check if t least two binners where applied in order to run DAS Tool for bin refinment + // (needs to be adjusted in case additional binners are added) + if (params.refine_bins_dastool && params.skip_metabat2 ){ + log.error "Both --refine_bins_dastool and --skip_metabat2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results." + System.exit(1) + } + if (params.refine_bins_dastool && params.skip_maxbin2 ){ + log.error "Both --refine_bins_dastool and --skip_maxbin2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results." + System.exit(1) + } + // Check if BUSCO parameters combinations are valid if (params.skip_busco){ if (params.busco_reference) { From 693f7c55ab22c5acc05453160a77954eb79ffc1b Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Thu, 14 Apr 2022 13:06:42 +0200 Subject: [PATCH 156/246] Update lib/WorkflowMag.groovy --- lib/WorkflowMag.groovy | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/lib/WorkflowMag.groovy b/lib/WorkflowMag.groovy index 33f4ab7b..70aed353 100755 --- a/lib/WorkflowMag.groovy +++ b/lib/WorkflowMag.groovy @@ -79,7 +79,7 @@ class WorkflowMag { } } - // Check if t least two binners where applied in order to run DAS Tool for bin refinment + // Check if at least two binners were applied in order to run DAS Tool for bin refinment // (needs to be adjusted in case additional binners are added) if (params.refine_bins_dastool && params.skip_metabat2 ){ log.error "Both --refine_bins_dastool and --skip_metabat2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results." From bba156ac1101c78e65541b62700161853630d002 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 14 Apr 2022 13:26:10 +0200 Subject: [PATCH 157/246] Move postbinning check to other checks change publishing option --- conf/modules.config | 12 ++++--- lib/WorkflowMag.groovy | 71 ++++++++++++++++++++++++------------------ workflows/mag.nf | 2 -- 3 files changed, 48 insertions(+), 37 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 937f80a4..31e85e1c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -457,17 +457,19 @@ process { mode: params.publish_dir_mode, pattern: '*.{tsv,log,eval,seqlength}' ], - [ - path: { "${params.outdir}/BinRefinement/" }, - mode: params.publish_dir_mode, - pattern: '*_bins/*.fa' - ], ] ext.prefix = { "${meta.assembler}-DASTool-${meta.id}" } ext.args = "--write_bins --write_unbinned --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" } withName: RENAME_DASTOOL { + publishDir = [ + [ + path: { "${params.outdir}/BinRefinement/" }, + mode: params.publish_dir_mode, + pattern: '*.fasta' + ], + ] ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } } diff --git a/lib/WorkflowMag.groovy b/lib/WorkflowMag.groovy index 70aed353..859e6fc6 100755 --- a/lib/WorkflowMag.groovy +++ b/lib/WorkflowMag.groovy @@ -9,7 +9,7 @@ class WorkflowMag { // public static void initialise(params, log, hybrid) { // Check if binning mapping mode is valid - if (!['all','group','own'].contains(params.binning_map_mode)) { + if (!['all', 'group', 'own'].contains(params.binning_map_mode)) { log.error "Invalid parameter '--binning_map_mode ${params.binning_map_mode}'. Valid values are 'all', 'group' or 'own'." System.exit(1) } @@ -28,45 +28,49 @@ class WorkflowMag { System.exit(1) } // Check if settings concerning reproducibility of used tools are consistent and print warning if not - if (params.megahit_fix_cpu_1 || params.spades_fix_cpus != -1 || params.spadeshybrid_fix_cpus != -1){ - if (!params.skip_spades && params.spades_fix_cpus == -1) + if (params.megahit_fix_cpu_1 || params.spades_fix_cpus != -1 || params.spadeshybrid_fix_cpus != -1) { + if (!params.skip_spades && params.spades_fix_cpus == -1) { log.warn "At least one assembly process is run with a parameter to ensure reproducible results, but SPAdes not. Consider using the parameter '--spades_fix_cpus'." - if (hybrid && params.skip_spadeshybrid && params.spadeshybrid_fix_cpus == -1) + } + if (hybrid && params.skip_spadeshybrid && params.spadeshybrid_fix_cpus == -1) { log.warn "At least one assembly process is run with a parameter to ensure reproducible results, but SPAdes hybrid not. Consider using the parameter '--spadeshybrid_fix_cpus'." - if (!params.skip_megahit && !params.megahit_fix_cpu_1) + } + if (!params.skip_megahit && !params.megahit_fix_cpu_1) { log.warn "At least one assembly process is run with a parameter to ensure reproducible results, but MEGAHIT not. Consider using the parameter '--megahit_fix_cpu_1'." - if (!params.skip_binning && params.metabat_rng_seed == 0) + } + if (!params.skip_binning && params.metabat_rng_seed == 0) { log.warn "At least one assembly process is run with a parameter to ensure reproducible results, but for MetaBAT2 a random seed is specified ('--metabat_rng_seed 0'). Consider specifying a positive seed instead." + } } // Check if SPAdes and single_end if ( (!params.skip_spades || !params.skip_spadeshybrid) && params.single_end) { - log.warn "metaSPAdes does not support single-end data. SPAdes will be skipped." + log.warn 'metaSPAdes does not support single-end data. SPAdes will be skipped.' } // Check if parameters for host contamination removal are valid if ( params.host_fasta && params.host_genome) { - log.error "Both host fasta reference and iGenomes genome are specified to remove host contamination! Invalid combination, please specify either --host_fasta or --host_genome." + log.error 'Both host fasta reference and iGenomes genome are specified to remove host contamination! Invalid combination, please specify either --host_fasta or --host_genome.' System.exit(1) } if ( hybrid && (params.host_fasta || params.host_genome) ) { - log.warn "Host read removal is only applied to short reads. Long reads might be filtered indirectly by Filtlong, which is set to use read qualities estimated based on k-mer matches to the short, already filtered reads." + log.warn 'Host read removal is only applied to short reads. Long reads might be filtered indirectly by Filtlong, which is set to use read qualities estimated based on k-mer matches to the short, already filtered reads.' if ( params.longreads_length_weight > 1 ) { log.warn "The parameter --longreads_length_weight is ${params.longreads_length_weight}, causing the read length being more important for long read filtering than the read quality. Set --longreads_length_weight to 1 in order to assign equal weights." } } if ( params.host_genome ) { if (!params.genomes) { - log.error "No config file containing genomes provided!" + log.error 'No config file containing genomes provided!' System.exit(1) } // Check if host genome exists in the config file if (!params.genomes.containsKey(params.host_genome)) { - log.error "=============================================================================\n" + + log.error '=============================================================================\n' + " Host genome '${params.host_genome}' not found in any config files provided to the pipeline.\n" + - " Currently, the available genome keys are:\n" + - " ${params.genomes.keySet().join(", ")}\n" + - "===================================================================================" + ' Currently, the available genome keys are:\n' + + " ${params.genomes.keySet().join(', ')}\n" + + '===================================================================================' System.exit(1) } if ( !params.genomes[params.host_genome].fasta ) { @@ -81,50 +85,56 @@ class WorkflowMag { // Check if at least two binners were applied in order to run DAS Tool for bin refinment // (needs to be adjusted in case additional binners are added) - if (params.refine_bins_dastool && params.skip_metabat2 ){ - log.error "Both --refine_bins_dastool and --skip_metabat2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results." + if (params.refine_bins_dastool && params.skip_metabat2 ) { + log.error 'Both --refine_bins_dastool and --skip_metabat2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results.' System.exit(1) } - if (params.refine_bins_dastool && params.skip_maxbin2 ){ - log.error "Both --refine_bins_dastool and --skip_maxbin2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results." + if (params.refine_bins_dastool && params.skip_maxbin2 ) { + log.error 'Both --refine_bins_dastool and --skip_maxbin2 are specified! Invalid combination, bin refinement requires MetaBAT2 and MaxBin2 binning results.' + System.exit(1) + } + + // Check that bin refinement is actually turned on if any of the refined bins are requested for downstream + if (!params.refine_bins_dastool && params.postbinning_input != 'raw_bins_only') { + log.error "[nf-core/mag] error: Can only specify '--postbinning_input ${params.postbinning_input}' for downstream steps if bin refinement activated with --refine_bins_dastool! Check input." System.exit(1) } // Check if BUSCO parameters combinations are valid - if (params.skip_busco){ + if (params.skip_busco) { if (params.busco_reference) { - log.error "Both --skip_busco and --busco_reference are specified! Invalid combination, please specify either --skip_busco or --busco_reference." + log.error 'Both --skip_busco and --busco_reference are specified! Invalid combination, please specify either --skip_busco or --busco_reference.' System.exit(1) } if (params.busco_download_path) { - log.error "Both --skip_busco and --busco_download_path are specified! Invalid combination, please specify either --skip_busco or --busco_download_path." + log.error 'Both --skip_busco and --busco_download_path are specified! Invalid combination, please specify either --skip_busco or --busco_download_path.' System.exit(1) } if (params.busco_auto_lineage_prok) { - log.error "Both --skip_busco and --busco_auto_lineage_prok are specified! Invalid combination, please specify either --skip_busco or --busco_auto_lineage_prok." + log.error 'Both --skip_busco and --busco_auto_lineage_prok are specified! Invalid combination, please specify either --skip_busco or --busco_auto_lineage_prok.' System.exit(1) } } if (params.busco_reference && params.busco_download_path) { - log.error "Both --busco_reference and --busco_download_path are specified! Invalid combination, please specify either --busco_reference or --busco_download_path." + log.error 'Both --busco_reference and --busco_download_path are specified! Invalid combination, please specify either --busco_reference or --busco_download_path.' System.exit(1) } if (params.busco_auto_lineage_prok && params.busco_reference) { - log.error "Both --busco_auto_lineage_prok and --busco_reference are specified! Invalid combination, please specify either --busco_auto_lineage_prok or --busco_reference." + log.error 'Both --busco_auto_lineage_prok and --busco_reference are specified! Invalid combination, please specify either --busco_auto_lineage_prok or --busco_reference.' System.exit(1) } if (params.skip_busco && params.gtdb) { - log.warn "--skip_busco and --gtdb are specified! GTDB-tk will be omitted because GTDB-tk bin classification requires bin filtering based on BUSCO QC results to avoid GTDB-tk errors." + log.warn '--skip_busco and --gtdb are specified! GTDB-tk will be omitted because GTDB-tk bin classification requires bin filtering based on BUSCO QC results to avoid GTDB-tk errors.' } // Check if CAT parameters are valid if (params.cat_db && params.cat_db_generate) { - log.error "Invalid combination of parameters --cat_db and --cat_db_generate is specified! Please specify either --cat_db or --cat_db_generate." + log.error 'Invalid combination of parameters --cat_db and --cat_db_generate is specified! Please specify either --cat_db or --cat_db_generate.' System.exit(1) } if (params.save_cat_db && !params.cat_db_generate) { - log.error "Invalid parameter combination: parameter --save_cat_db specified, but not --cat_db_generate! Note also that the parameter --save_cat_db does not work in combination with --cat_db." + log.error 'Invalid parameter combination: parameter --save_cat_db specified, but not --cat_db_generate! Note also that the parameter --save_cat_db does not work in combination with --cat_db.' System.exit(1) } } @@ -142,17 +152,18 @@ class WorkflowMag { for (param in group_params.keySet()) { summary_section += "
$param
${group_params.get(param) ?: 'N/A'}
\n" } - summary_section += " \n" + summary_section += ' \n' } } - String yaml_file_text = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n" + String yaml_file_text = "id: '${workflow.manifest.name.replace('/', '-')}-summary'\n" yaml_file_text += "description: ' - this information is collected when the pipeline is started.'\n" yaml_file_text += "section_name: '${workflow.manifest.name} Workflow Summary'\n" yaml_file_text += "section_href: 'https://github.com/${workflow.manifest.name}'\n" yaml_file_text += "plot_type: 'html'\n" - yaml_file_text += "data: |\n" + yaml_file_text += 'data: |\n' yaml_file_text += "${summary_section}" return yaml_file_text } + } diff --git a/workflows/mag.nf b/workflows/mag.nf index db6df316..c9b85610 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -35,8 +35,6 @@ for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true // Check mandatory parameters if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input samplesheet not specified!' } -if ( !params.refine_bins_dastool && params.postbinning_input != 'raw_bins_only' ) { exit 1, "[nf-core/mag] error: Can only specify '--postbinning_input ${params.postbinning_input}' for downstream steps if bin refinement activated with --refine_bins_dastool! Check input." } - /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ CONFIG FILES From ff32e18b7bdd53afebc9193210af90aa47bb8391 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 14 Apr 2022 15:33:30 +0200 Subject: [PATCH 158/246] Attempt to pre-rename input bins to DAS_Tool to ensure these have corect downstream --- ...ename_dastool.nf => rename_postdastool.nf} | 8 ++---- modules/local/rename_predastool.nf | 26 +++++++++++++++++++ subworkflows/local/binning_refinement.nf | 19 +++++++------- 3 files changed, 38 insertions(+), 15 deletions(-) rename modules/local/{rename_dastool.nf => rename_postdastool.nf} (83%) create mode 100644 modules/local/rename_predastool.nf diff --git a/modules/local/rename_dastool.nf b/modules/local/rename_postdastool.nf similarity index 83% rename from modules/local/rename_dastool.nf rename to modules/local/rename_postdastool.nf index 3f7bf6cb..3a7505ec 100644 --- a/modules/local/rename_dastool.nf +++ b/modules/local/rename_postdastool.nf @@ -1,4 +1,4 @@ -process RENAME_DASTOOL { +process RENAME_POSTDASTOOL { tag "${meta.assembler}-${meta.id}" label 'process_low' @@ -18,11 +18,7 @@ process RENAME_DASTOOL { script: """ if [[ -f unbinned.fa ]]; then - mv unbinned.fa ${meta.assembler}-${meta.binner}-${meta.id}_refined_unbinned.fasta + mv unbinned.fa ${meta.assembler}-DASToolRefined-${meta.id}_refined_unbinned.fasta fi - - for i in *.fa; do - mv \$i \${i%%.fa}_refined.fasta - done """ } diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf new file mode 100644 index 00000000..4167aa06 --- /dev/null +++ b/modules/local/rename_predastool.nf @@ -0,0 +1,26 @@ +process RENAME_PREDASTOOL { + tag "${meta.assembler}-${meta.id}" + label 'process_low' + + // Using container from multiqc since it'll be included anyway + conda (params.enable_conda ? "bioconda::multiqc=1.12" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.12--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.12--pyhdfd78af_0' }" + + input: + tuple val(meta), path(bins) + + output: + tuple val(meta), path("${meta.assembler}-${meta.binner}Refined-${meta.id}*") , optional:true, emit: renamed_bins + + script: + """ + for i in *{fa.gz,fasta.gz}; do + filename=\$i + suffix=\${filename#*.} + + mv \$i ${meta.assembler}-${meta.binner}Refined-${meta.id}.\$suffix + done + """ +} diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index c6074bdf..a4a715db 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -5,7 +5,8 @@ include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_METABAT2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 } from '../../modules/nf-core/modules/dastool/fastatocontig2bin/main.nf' include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' -include { RENAME_DASTOOL } from '../../modules/local/rename_dastool' +include { RENAME_PREDASTOOL } from '../../modules/local/rename_predastool' +include { RENAME_POSTDASTOOL } from '../../modules/local/rename_postdastool' workflow BINNING_REFINEMENT { @@ -23,11 +24,11 @@ workflow BINNING_REFINEMENT { [ meta_new, assembly ] } - ch_bins_for_fastatocontig2bin = bins - .branch { - metabat2: it[0]['binner'] == 'MetaBAT2' - maxbin2: it[0]['binner'] == 'MaxBin2' - } + ch_bins_for_fastatocontig2bin = RENAME_PREDASTOOL(bins).renamed_bins + .branch { + metabat2: it[0]['binner'] == 'MetaBAT2' + maxbin2: it[0]['binner'] == 'MaxBin2' + } // Run on each bin file separately DASTOOL_FASTATOCONTIG2BIN_METABAT2 ( ch_bins_for_fastatocontig2bin.metabat2, "fa") @@ -62,10 +63,10 @@ workflow BINNING_REFINEMENT { [ meta_new, bins ] } - RENAME_DASTOOL ( ch_input_for_renamedastool ) + RENAME_POSTDASTOOL ( ch_input_for_renamedastool ) emit: - refined_bins = RENAME_DASTOOL.out.refined_bins - refined_unbins = RENAME_DASTOOL.out.refined_unbins + refined_bins = DASTOOL_DASTOOL.out.bins + refined_unbins = RENAME_POSTDASTOOL.out.refined_unbins versions = ch_versions } From 9136428aaeb4738d4b0e27a2ac9e8da80dd07544 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 14 Apr 2022 15:34:19 +0200 Subject: [PATCH 159/246] Bash fix --- modules/local/rename_predastool.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index 4167aa06..f6564580 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -16,7 +16,7 @@ process RENAME_PREDASTOOL { script: """ - for i in *{fa.gz,fasta.gz}; do + for i in ${bins}; do filename=\$i suffix=\${filename#*.} From dc92886209927143dae8ef3daa0d5ad5dcf5a6ce Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 19 Apr 2022 12:11:42 +0200 Subject: [PATCH 160/246] Add error handling in BUSCO_PLOT --- modules/local/busco_plot.nf | 16 ++++++++++------ 1 file changed, 10 insertions(+), 6 deletions(-) diff --git a/modules/local/busco_plot.nf b/modules/local/busco_plot.nf index ecda860c..26abf6a6 100644 --- a/modules/local/busco_plot.nf +++ b/modules/local/busco_plot.nf @@ -21,12 +21,16 @@ process BUSCO_PLOT { # replace dots in bin names within summary file names by underscores # currently (BUSCO v5.1.0) generate_plot.py does not allow further dots for sum in ${summaries}; do - [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt ]]; - mode=\${BASH_REMATCH[1]} - db_name=\${BASH_REMATCH[2]} - bin="${meta.assembler}-${meta.binner}-${meta.id}.\${BASH_REMATCH[3]}" - bin_new="\${bin//./_}" - mv \${sum} short_summary.\${mode}.\${db_name}.\${bin_new}.txt + if [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt ]]; then + mode=\${BASH_REMATCH[1]} + db_name=\${BASH_REMATCH[2]} + bin="${meta.assembler}-${meta.binner}-${meta.id}.\${BASH_REMATCH[3]}" + bin_new="\${bin//./_}" + mv \${sum} short_summary.\${mode}.\${db_name}.\${bin_new}.txt + else + echo "ERROR: the summary filename \${sum} does not match the expected format 'short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt'!" + exit 1 + fi done generate_plot.py --working_directory . From 378f139a87169e90761b9ce9fcf8cbb86df97009 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 19 Apr 2022 12:12:42 +0200 Subject: [PATCH 161/246] Fix output filenames in BUSCO_PLOT --- modules/local/busco_plot.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/modules/local/busco_plot.nf b/modules/local/busco_plot.nf index 26abf6a6..c3ff6a8e 100644 --- a/modules/local/busco_plot.nf +++ b/modules/local/busco_plot.nf @@ -1,5 +1,5 @@ process BUSCO_PLOT { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" conda (params.enable_conda ? "bioconda::busco=5.1.0" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -10,8 +10,8 @@ process BUSCO_PLOT { tuple val(meta), path(summaries) output: - path("${meta.assembler}-${meta.id}.*.busco_figure.png") , optional:true, emit: png - path("${meta.assembler}-${meta.id}.*.busco_figure.R") , optional:true, emit: rscript + path("${meta.assembler}-${meta.binner}-${meta.id}.*.busco_figure.png") , optional:true, emit: png + path("${meta.assembler}-${meta.binner}-${meta.id}.*.busco_figure.R") , optional:true, emit: rscript path "versions.yml" , emit: versions script: From e07521cde983c976f5f1a1c3bb11c57887ccaf7b Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 28 Apr 2022 15:32:02 +0200 Subject: [PATCH 162/246] Fix BUSCO issue --- conf/modules.config | 15 ++++++- modules/local/rename_postdastool.nf | 5 +-- modules/local/rename_predastool.nf | 7 ++- subworkflows/local/binning.nf | 3 +- subworkflows/local/binning_refinement.nf | 56 +++++++++++++++++++++--- workflows/mag.nf | 6 ++- 6 files changed, 75 insertions(+), 17 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 31e85e1c..28048a8f 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -462,12 +462,23 @@ process { ext.args = "--write_bins --write_unbinned --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" } - withName: RENAME_DASTOOL { + withName: RENAME_PREDASTOOL { publishDir = [ [ path: { "${params.outdir}/BinRefinement/" }, mode: params.publish_dir_mode, - pattern: '*.fasta' + pattern: '*.fa' + ], + ] + ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } + } + + withName: RENAME_POSTDASTOOL { + publishDir = [ + [ + path: { "${params.outdir}/BinRefinement/" }, + mode: params.publish_dir_mode, + pattern: '*.fa' ], ] ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } diff --git a/modules/local/rename_postdastool.nf b/modules/local/rename_postdastool.nf index 3a7505ec..8691b525 100644 --- a/modules/local/rename_postdastool.nf +++ b/modules/local/rename_postdastool.nf @@ -12,13 +12,12 @@ process RENAME_POSTDASTOOL { tuple val(meta), path(bins) output: - tuple val(meta), path("*_refined.fasta") , optional:true, emit: refined_bins - tuple val(meta), path("*_refined_unbinned.fasta") , optional:true, emit: refined_unbins + tuple val(meta), path("${meta.assembler}-DASToolUnbinned-${meta.id}.fa") , optional:true, emit: refined_unbins script: """ if [[ -f unbinned.fa ]]; then - mv unbinned.fa ${meta.assembler}-DASToolRefined-${meta.id}_refined_unbinned.fasta + mv unbinned.fa ${meta.assembler}-DASToolUnbinned-${meta.id}.fa fi """ } diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index f6564580..5e3a4e2e 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -16,11 +16,10 @@ process RENAME_PREDASTOOL { script: """ - for i in ${bins}; do - filename=\$i - suffix=\${filename#*.} + SAMPLES=(*) - mv \$i ${meta.assembler}-${meta.binner}Refined-${meta.id}.\$suffix + for i in \$(seq 0 \$((\${#SAMPLES[@]}-1))); do + mv \${SAMPLES[\$i]} ${meta.assembler}-${meta.binner}Refined-${meta.id}.\$(( \$i + 1)).fa done """ } diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 9805d450..6dd5b117 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -67,7 +67,7 @@ workflow BINNING { [ meta, assembly, depths ] } - // conver metabat2 depth files to maxbin2 + // convert metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) ch_maxbin2_input = CONVERT_DEPTHS.out.output @@ -179,5 +179,6 @@ workflow BINNING { unbinned = ch_splitfasta_results_gunzipped.groupTuple() unbinned_gz = SPLIT_FASTA.out.unbinned depths_summary = MAG_DEPTHS_SUMMARY.out.summary + metabat2depths = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth versions = ch_versions } diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index a4a715db..a5d318f6 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -7,16 +7,18 @@ include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 } from include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' include { RENAME_PREDASTOOL } from '../../modules/local/rename_predastool' include { RENAME_POSTDASTOOL } from '../../modules/local/rename_postdastool' - +include { MAG_DEPTHS } from '../../modules/local/mag_depths' workflow BINNING_REFINEMENT { take: contigs // bins // channel: [ val(meta), path(bins) ] + depths main: ch_versions = Channel.empty() + // Drop unnecessary files ch_contigs_for_dastool = contigs .map { meta, assembly, bams, bais -> @@ -24,16 +26,25 @@ workflow BINNING_REFINEMENT { [ meta_new, assembly ] } + // Rename for consistency for downstream steps (BUSCO etc.) ch_bins_for_fastatocontig2bin = RENAME_PREDASTOOL(bins).renamed_bins + .map { + meta, bins -> + def meta_new = meta.clone() + meta_new.binner = meta.binner + 'Refined' + + [ meta_new, bins ] + } .branch { - metabat2: it[0]['binner'] == 'MetaBAT2' - maxbin2: it[0]['binner'] == 'MaxBin2' + metabat2: it[0]['binner'] == 'MetaBAT2Refined' + maxbin2: it[0]['binner'] == 'MaxBin2Refined' } - // Run on each bin file separately + // Generate DASTool auxilary files DASTOOL_FASTATOCONTIG2BIN_METABAT2 ( ch_bins_for_fastatocontig2bin.metabat2, "fa") DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 ( ch_bins_for_fastatocontig2bin.maxbin2, "fasta") + // Ru DAST ch_fastatocontig2bin_for_dastool = Channel.empty() ch_fastatocontig2bin_for_dastool = ch_fastatocontig2bin_for_dastool .mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.fastatocontig2bin) @@ -46,13 +57,24 @@ workflow BINNING_REFINEMENT { } .groupTuple(by: 0) - ch_input_for_dastool = ch_contigs_for_dastool.join(ch_fastatocontig2bin_for_dastool, by: 0).dump(tag: "input_to_dastool") + ch_input_for_dastool = ch_contigs_for_dastool.join(ch_fastatocontig2bin_for_dastool, by: 0) ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.versions.first()) ch_versions = ch_versions.mix(DASTOOL_FASTATOCONTIG2BIN_MAXBIN2.out.versions.first()) // Run DAStool DASTOOL_DASTOOL(ch_input_for_dastool, [], []) + + ch_dastool_bins_newmeta = DASTOOL_DASTOOL.out.bins.transpose() + .map { + meta, bins -> + def meta_new = meta.clone() + + meta_new['binner'] = bins.name.split("-")[1] + [ meta_new, bins ] + } + .groupTuple() + ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions.first()) ch_input_for_renamedastool = DASTOOL_DASTOOL.out.bins @@ -65,8 +87,30 @@ workflow BINNING_REFINEMENT { RENAME_POSTDASTOOL ( ch_input_for_renamedastool ) + // We have to strip the meta to be able to combine with the original + // depths file to run MAG_DEPTH + depths.dump(tag: "depths") + + ch_input_for_magdepth = ch_dastool_bins_newmeta + .mix( RENAME_POSTDASTOOL.out.refined_unbins ) + .map { + meta, refinedbins -> + def meta_new = [ 'id': meta.id, 'group': meta.group, 'single_end': meta.single_end, 'assembler': meta.assembler ] + [ meta_new, refinedbins ] + } + .join( depths, by: 0 ) + .dump(tag: "refined_cominbed_join") + + MAG_DEPTHS ( ch_input_for_magdepth ) + + // TODO Add MAG_DEPTH_SUMMARY + // Update channel logic in mag.nf for BIN_SUMARY so if refined use bin refined DPETH_SUMMARY, otherwise use original from binning.nf + // Work out what to do when running refined + raw together? Separate plots? + emit: - refined_bins = DASTOOL_DASTOOL.out.bins + refined_bins = ch_dastool_bins_newmeta refined_unbins = RENAME_POSTDASTOOL.out.refined_unbins + refined_bin_depths = MAG_DEPTHS.out.depths versions = ch_versions } + diff --git a/workflows/mag.nf b/workflows/mag.nf index c9b85610..632d52d9 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -553,7 +553,8 @@ workflow MAG { * DAS_Tool: binning refinement */ if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { - BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins ) + + BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins, BINNING.out.metabat2depths ) ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) if ( params.postbinning_input == 'raw_bins_only' ) { @@ -572,6 +573,8 @@ workflow MAG { ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) } + ch_input_for_postbinning_bins + ch_input_for_postbinning_bins if (!params.skip_busco){ /* @@ -639,6 +642,7 @@ workflow MAG { ch_gtdbtk_summary = GTDBTK.out.summary } + if (!params.skip_busco || !params.skip_quast || gtdb){ BIN_SUMMARY ( BINNING.out.depths_summary, From 98a29a278c99befaa6ef4e0621d141f9fddd6728 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 29 Apr 2022 10:35:26 +0200 Subject: [PATCH 163/246] Undo .set{} replacements in `mag.nf` to do in another PR --- subworkflows/local/binning.nf | 15 ++++++++++----- workflows/mag.nf | 7 ++++--- 2 files changed, 14 insertions(+), 8 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 6dd5b117..324696b9 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -121,17 +121,19 @@ workflow BINNING { .mix( ch_maxbin2_results_transposed ) GUNZIP_BINS ( ch_final_bins_for_gunzip ) - ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip + GUNZIP_BINS.out + .set{ ch_binning_results_gunzipped } ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions.first()) GUNZIP_UNBINS ( ch_split_fasta_results_transposed ) - ch_splitfasta_results_gunzipped = GUNZIP_UNBINS.out.gunzip + GUNZIP_UNBINS.out.gunzip + .set{ ch_splitfasta_results_gunzipped } ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions.first()) // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta for grouping to mix back with original depth // files, as required for MAG_DEPTHS - ch_depth_input = ch_binning_results_gunzipped + ch_binning_results_gunzipped .mix(ch_splitfasta_results_gunzipped ) .map { meta, results -> def meta_new = meta.clone() @@ -140,6 +142,7 @@ workflow BINNING { } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) @@ -165,11 +168,13 @@ workflow BINNING { ch_versions = ch_versions.mix( MAG_DEPTHS_SUMMARY.out.versions ) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped_final = ch_binning_results_gunzipped + ch_binning_results_gunzipped .groupTuple(by: 0) + .set{ ch_binning_results_gunzipped_final } - ch_binning_results_gzipped_final = METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) + METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) .groupTuple(by: 0) + .set{ ch_binning_results_gzipped_final } SPLIT_FASTA.out.unbinned diff --git a/workflows/mag.nf b/workflows/mag.nf index 632d52d9..80a03128 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -314,13 +314,15 @@ workflow MAG { } // join long and short reads by sample name - ch_short_reads_tmp = ch_short_reads + ch_short_reads .map { meta, sr -> [ meta.id, meta, sr ] } + .set { ch_short_reads_tmp } - ch_short_and_long_reads = ch_long_reads + ch_long_reads .map { meta, lr -> [ meta.id, meta, lr ] } .join(ch_short_reads_tmp, by: 0) .map { id, meta_lr, lr, meta_sr, sr -> [ meta_lr, lr, sr[0], sr[1] ] } // should not occur for single-end, since SPAdes (hybrid) does not support single-end + .set { ch_short_and_long_reads } FILTLONG ( ch_short_and_long_reads @@ -642,7 +644,6 @@ workflow MAG { ch_gtdbtk_summary = GTDBTK.out.summary } - if (!params.skip_busco || !params.skip_quast || gtdb){ BIN_SUMMARY ( BINNING.out.depths_summary, From 0fce6b34f0c461a133c4dd8b0cfc7195078eb800 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 29 Apr 2022 10:38:02 +0200 Subject: [PATCH 164/246] Revert formatting --- subworkflows/local/binning.nf | 2 +- workflows/mag.nf | 4 ++-- 2 files changed, 3 insertions(+), 3 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 324696b9..1788cbb9 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -121,7 +121,7 @@ workflow BINNING { .mix( ch_maxbin2_results_transposed ) GUNZIP_BINS ( ch_final_bins_for_gunzip ) - GUNZIP_BINS.out + GUNZIP_BINS.out.gunzip .set{ ch_binning_results_gunzipped } ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions.first()) diff --git a/workflows/mag.nf b/workflows/mag.nf index 80a03128..a3a50bcd 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -316,13 +316,13 @@ workflow MAG { // join long and short reads by sample name ch_short_reads .map { meta, sr -> [ meta.id, meta, sr ] } - .set { ch_short_reads_tmp } + .set {ch_short_reads_tmp} ch_long_reads .map { meta, lr -> [ meta.id, meta, lr ] } .join(ch_short_reads_tmp, by: 0) .map { id, meta_lr, lr, meta_sr, sr -> [ meta_lr, lr, sr[0], sr[1] ] } // should not occur for single-end, since SPAdes (hybrid) does not support single-end - .set { ch_short_and_long_reads } + .set{ ch_short_and_long_reads } FILTLONG ( ch_short_and_long_reads From 97763c524ef707a38ef8972c8728aa272abc0f3c Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 14:41:38 +0200 Subject: [PATCH 165/246] Finalise bin_summary update --- conf/modules.config | 31 ++++++++++++++++-- modules/local/combine_tsv.nf | 26 +++++++++++++++ modules/local/mag_depths_summary.nf | 5 +-- subworkflows/local/binning_refinement.nf | 41 +++++++++++++++++------- workflows/mag.nf | 18 ++++++++--- 5 files changed, 99 insertions(+), 22 deletions(-) create mode 100644 modules/local/combine_tsv.nf diff --git a/conf/modules.config b/conf/modules.config index 28048a8f..708caac6 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -217,6 +217,31 @@ process { ] } + withName: 'BIN_SUMMARY' { + publishDir = [ + path: params.refine_bins_dastool ? { "${params.outdir}/GenomeBinningRefinement" } : { "${params.outdir}/GenomeBinning" }, + mode: params.publish_dir_mode, + pattern: "*.{png,tsv}" + ] + } + + withName: 'MAG_DEPTHS_PLOT_REFINED|BIN_SUMMARY_REFINED' { + publishDir = [ + path: { "${params.outdir}/GenomeBinningRefinement/" }, + mode: params.publish_dir_mode, + pattern: "*.{png,tsv}" + ] + } + + withName: 'MAG_DEPTHS_SUMMARY_REFINED' { + ext.prefix = "bin_depths_summary_refined" + publishDir = [ + path: { "${params.outdir}/GenomeBinningRefinement/" }, + mode: params.publish_dir_mode, + pattern: "*.{tsv}" + ] + } + withName: BUSCO_DB_PREPARATION { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, @@ -453,7 +478,7 @@ process { withName: DASTOOL_DASTOOL { publishDir = [ [ - path: { "${params.outdir}/BinRefinement/" }, + path: { "${params.outdir}/GenomeBinRefinement/" }, mode: params.publish_dir_mode, pattern: '*.{tsv,log,eval,seqlength}' ], @@ -465,7 +490,7 @@ process { withName: RENAME_PREDASTOOL { publishDir = [ [ - path: { "${params.outdir}/BinRefinement/" }, + path: { "${params.outdir}/GenomeBinRefinement/" }, mode: params.publish_dir_mode, pattern: '*.fa' ], @@ -476,7 +501,7 @@ process { withName: RENAME_POSTDASTOOL { publishDir = [ [ - path: { "${params.outdir}/BinRefinement/" }, + path: { "${params.outdir}/GenomeBinRefinement/" }, mode: params.publish_dir_mode, pattern: '*.fa' ], diff --git a/modules/local/combine_tsv.nf b/modules/local/combine_tsv.nf new file mode 100644 index 00000000..789e4856 --- /dev/null +++ b/modules/local/combine_tsv.nf @@ -0,0 +1,26 @@ +process COMBINE_TSV { + + // Using bioawk as already use that for CONVERT_DEPTHS and does same thing + conda (params.enable_conda ? "bioconda::bioawk=1.0" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bioawk:1.0--hed695b0_5' : + 'quay.io/biocontainers/bioawk:1.0--hed695b0_5' }" + + input: + path(bin_summaries) + + output: + path("*.tsv"), emit: combined + path "versions.yml" , emit: versions + + script: + def prefix = task.ext.prefix ?: "bin_depths_summary_combined" + """ + bioawk '(NR == 1) || (FNR > 1)' ${bin_summaries} > ${prefix}.tsv + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bioawk: \$(bioawk --version | cut -f 3 -d ' ' ) + END_VERSIONS + """ +} diff --git a/modules/local/mag_depths_summary.nf b/modules/local/mag_depths_summary.nf index 95a8ffde..a3c9e43f 100644 --- a/modules/local/mag_depths_summary.nf +++ b/modules/local/mag_depths_summary.nf @@ -9,13 +9,14 @@ process MAG_DEPTHS_SUMMARY { path(mag_depths) output: - path("bin_depths_summary.tsv"), emit: summary + path("${prefix}.tsv"), emit: summary path "versions.yml" , emit: versions script: + prefix = task.ext.prefix ?: "bin_depths_summary" """ get_mag_depths_summary.py --depths ${mag_depths} \ - --out "bin_depths_summary.tsv" + --out "${prefix}.tsv" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index a5d318f6..7f04d5fd 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -7,13 +7,24 @@ include { DASTOOL_FASTATOCONTIG2BIN as DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 } from include { DASTOOL_DASTOOL } from '../../modules/nf-core/modules/dastool/dastool/main.nf' include { RENAME_PREDASTOOL } from '../../modules/local/rename_predastool' include { RENAME_POSTDASTOOL } from '../../modules/local/rename_postdastool' -include { MAG_DEPTHS } from '../../modules/local/mag_depths' +include { MAG_DEPTHS as MAG_DEPTHS_REFINED } from '../../modules/local/mag_depths' +include { MAG_DEPTHS_PLOT as MAG_DEPTHS_PLOT_REFINED } from '../../modules/local/mag_depths_plot' +include { MAG_DEPTHS_SUMMARY as MAG_DEPTHS_SUMMARY_REFINED } from '../../modules/local/mag_depths_summary' + +/* + * Get number of columns in file (first line) + */ +def getColNo(filename) { + lines = file(filename).readLines() + return lines[0].split('\t').size() +} workflow BINNING_REFINEMENT { take: contigs // bins // channel: [ val(meta), path(bins) ] depths + reads main: ch_versions = Channel.empty() @@ -89,8 +100,6 @@ workflow BINNING_REFINEMENT { // We have to strip the meta to be able to combine with the original // depths file to run MAG_DEPTH - depths.dump(tag: "depths") - ch_input_for_magdepth = ch_dastool_bins_newmeta .mix( RENAME_POSTDASTOOL.out.refined_unbins ) .map { @@ -101,16 +110,24 @@ workflow BINNING_REFINEMENT { .join( depths, by: 0 ) .dump(tag: "refined_cominbed_join") - MAG_DEPTHS ( ch_input_for_magdepth ) + MAG_DEPTHS_REFINED ( ch_input_for_magdepth ) + + // Plot bin depths heatmap for each assembly and mapped samples (according to `binning_map_mode`) + // create file containing group information for all samples + ch_sample_groups = reads + .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } - // TODO Add MAG_DEPTH_SUMMARY - // Update channel logic in mag.nf for BIN_SUMARY so if refined use bin refined DPETH_SUMMARY, otherwise use original from binning.nf - // Work out what to do when running refined + raw together? Separate plots? + // filter MAG depth files: use only those for plotting that contain depths for > 2 samples + ch_mag_depths_plot_refined = MAG_DEPTHS_REFINED.out.depths + .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } + + MAG_DEPTHS_PLOT_REFINED ( ch_mag_depths_plot_refined, ch_sample_groups.collect() ) + MAG_DEPTHS_SUMMARY_REFINED ( MAG_DEPTHS_REFINED.out.depths.map{it[1]}.collect() ) emit: - refined_bins = ch_dastool_bins_newmeta - refined_unbins = RENAME_POSTDASTOOL.out.refined_unbins - refined_bin_depths = MAG_DEPTHS.out.depths - versions = ch_versions + refined_bins = ch_dastool_bins_newmeta + refined_unbins = RENAME_POSTDASTOOL.out.refined_unbins + refined_depths = MAG_DEPTHS_REFINED.out.depths + refined_depths_summary = MAG_DEPTHS_SUMMARY_REFINED.out.summary + versions = ch_versions } - diff --git a/workflows/mag.nf b/workflows/mag.nf index a3a50bcd..a4176e07 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -81,6 +81,7 @@ include { CAT_DB } from '../modules include { CAT_DB_GENERATE } from '../modules/local/cat_db_generate' include { CAT } from '../modules/local/cat' include { BIN_SUMMARY } from '../modules/local/bin_summary' +include { COMBINE_TSV } from '../modules/local/combine_tsv' include { MULTIQC } from '../modules/local/multiqc' // @@ -554,20 +555,30 @@ workflow MAG { /* * DAS_Tool: binning refinement */ + + // TODO Add MAG_DEPTH_SUMMARY + // Update channel logic in mag.nf for BIN_SUMARY so if refined use bin refined DPETH_SUMMARY, otherwise use original from binning.nf + // Work out what to do when running refined + raw together? Separate plots? + if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { - BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins, BINNING.out.metabat2depths ) + BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins, BINNING.out.metabat2depths, ch_short_reads ) ch_versions = ch_versions.mix(BINNING_REFINEMENT.out.versions) if ( params.postbinning_input == 'raw_bins_only' ) { ch_input_for_postbinning_bins = BINNING.out.bins ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) + ch_input_for_binsummary = BINNING.out.depths_summary } else if ( params.postbinning_input == 'refined_bins_only' ) { ch_input_for_postbinning_bins = BINNING_REFINEMENT.out.refined_bins ch_input_for_postbinning_bins_unbins = BINNING_REFINEMENT.out.refined_bins.mix(BINNING_REFINEMENT.out.refined_unbins) + ch_input_for_binsummary = BINNING_REFINEMENT.out.refined_depths_summary } else if (params.postbinning_input == 'both') { ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) + //TODO BINSUMMARY COMBINNING BOTH RAW AND REFINED + ch_combinedepthtsvs_for_binsummary = BINNING.out.depths_summary.mix(BINNING_REFINEMENT.out.refined_depths_summary).dump(tag: "input_to_combinetsv") + ch_input_for_binsummary = COMBINE_TSV ( ch_combinedepthtsvs_for_binsummary.collect() ).combined } } else { @@ -575,9 +586,6 @@ workflow MAG { ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) } - ch_input_for_postbinning_bins - ch_input_for_postbinning_bins - if (!params.skip_busco){ /* * BUSCO subworkflow: Quantitative measures for the assessment of genome assembly @@ -646,7 +654,7 @@ workflow MAG { if (!params.skip_busco || !params.skip_quast || gtdb){ BIN_SUMMARY ( - BINNING.out.depths_summary, + ch_input_for_binsummary, ch_busco_summary.ifEmpty([]), ch_quast_bins_summary.ifEmpty([]), ch_gtdbtk_summary.ifEmpty([]) From 698d753fddb45f7dbdf5f98c58fe8388f0d59913 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 14:43:23 +0200 Subject: [PATCH 166/246] Remove debugging --- conf/modules.config | 3 +++ subworkflows/local/binning_refinement.nf | 1 - workflows/mag.nf | 7 +------ 3 files changed, 4 insertions(+), 7 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 708caac6..b8bb2555 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -217,6 +217,9 @@ process { ] } + // Always push the publishDir for this step to the further downstream as + // possible, as it doesn't make sense to included refined bins in just the + // GenomeBinning directory withName: 'BIN_SUMMARY' { publishDir = [ path: params.refine_bins_dastool ? { "${params.outdir}/GenomeBinningRefinement" } : { "${params.outdir}/GenomeBinning" }, diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 7f04d5fd..7ec6ef72 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -108,7 +108,6 @@ workflow BINNING_REFINEMENT { [ meta_new, refinedbins ] } .join( depths, by: 0 ) - .dump(tag: "refined_cominbed_join") MAG_DEPTHS_REFINED ( ch_input_for_magdepth ) diff --git a/workflows/mag.nf b/workflows/mag.nf index a4176e07..e59d7460 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -556,10 +556,6 @@ workflow MAG { * DAS_Tool: binning refinement */ - // TODO Add MAG_DEPTH_SUMMARY - // Update channel logic in mag.nf for BIN_SUMARY so if refined use bin refined DPETH_SUMMARY, otherwise use original from binning.nf - // Work out what to do when running refined + raw together? Separate plots? - if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { BINNING_REFINEMENT ( BINNING_PREPARATION.out.grouped_mappings, BINNING.out.bins, BINNING.out.metabat2depths, ch_short_reads ) @@ -576,8 +572,7 @@ workflow MAG { } else if (params.postbinning_input == 'both') { ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) - //TODO BINSUMMARY COMBINNING BOTH RAW AND REFINED - ch_combinedepthtsvs_for_binsummary = BINNING.out.depths_summary.mix(BINNING_REFINEMENT.out.refined_depths_summary).dump(tag: "input_to_combinetsv") + ch_combinedepthtsvs_for_binsummary = BINNING.out.depths_summary.mix(BINNING_REFINEMENT.out.refined_depths_summary) ch_input_for_binsummary = COMBINE_TSV ( ch_combinedepthtsvs_for_binsummary.collect() ).combined } From 30348f1329372f996a9818b05c8c5b94d19e94bd Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Wed, 4 May 2022 14:48:38 +0200 Subject: [PATCH 167/246] Apply suggestions from code review Co-authored-by: Sabrina Krakau --- lib/WorkflowMag.groovy | 2 +- modules/local/rename_predastool.nf | 2 +- subworkflows/local/binning_refinement.nf | 8 ++++---- 3 files changed, 6 insertions(+), 6 deletions(-) diff --git a/lib/WorkflowMag.groovy b/lib/WorkflowMag.groovy index 859e6fc6..082027d6 100755 --- a/lib/WorkflowMag.groovy +++ b/lib/WorkflowMag.groovy @@ -96,7 +96,7 @@ class WorkflowMag { // Check that bin refinement is actually turned on if any of the refined bins are requested for downstream if (!params.refine_bins_dastool && params.postbinning_input != 'raw_bins_only') { - log.error "[nf-core/mag] error: Can only specify '--postbinning_input ${params.postbinning_input}' for downstream steps if bin refinement activated with --refine_bins_dastool! Check input." + log.error 'The parameter '--postbinning_input ${params.postbinning_input}' for downstream steps can only be specified if bin refinement is activated with --refine_bins_dastool! Check input.' System.exit(1) } diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index 5e3a4e2e..d25cf902 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -12,7 +12,7 @@ process RENAME_PREDASTOOL { tuple val(meta), path(bins) output: - tuple val(meta), path("${meta.assembler}-${meta.binner}Refined-${meta.id}*") , optional:true, emit: renamed_bins + tuple val(meta), path("${meta.assembler}-${meta.binner}Refined-${meta.id}*") , emit: renamed_bins script: """ diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 7ec6ef72..75163538 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -55,7 +55,7 @@ workflow BINNING_REFINEMENT { DASTOOL_FASTATOCONTIG2BIN_METABAT2 ( ch_bins_for_fastatocontig2bin.metabat2, "fa") DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 ( ch_bins_for_fastatocontig2bin.maxbin2, "fasta") - // Ru DAST + // Run DASTOOL ch_fastatocontig2bin_for_dastool = Channel.empty() ch_fastatocontig2bin_for_dastool = ch_fastatocontig2bin_for_dastool .mix(DASTOOL_FASTATOCONTIG2BIN_METABAT2.out.fastatocontig2bin) @@ -78,11 +78,11 @@ workflow BINNING_REFINEMENT { ch_dastool_bins_newmeta = DASTOOL_DASTOOL.out.bins.transpose() .map { - meta, bins -> + meta, bin -> def meta_new = meta.clone() - meta_new['binner'] = bins.name.split("-")[1] - [ meta_new, bins ] + meta_new['binner'] = bin.name.split("-")[1] + [ meta_new, bin ] } .groupTuple() From fc5f48e8c0b00f0ae6c8cf64177f750eac31d0f2 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 15:06:28 +0200 Subject: [PATCH 168/246] Revert "Add missing channel assignment" This reverts commit 1b9a20bc2bcea368eb3cbb7abde8ea7dd0e816a6. --- workflows/mag.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/mag.nf b/workflows/mag.nf index e59d7460..3f37d4f9 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -659,7 +659,7 @@ workflow MAG { /* * Prokka: Genome annotation */ - ch_bins_for_prokka = ch_input_for_postbinning_bins_unbins.transpose() + ch_input_for_postbinning_bins_unbins.transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() From 49a23da711774b8a1b42af887615f84a3bbdacc8 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 15:06:34 +0200 Subject: [PATCH 169/246] Revert "Remove duplicate channel assignment" This reverts commit 473893cfbf2c8d13ae233b74e53fbc6155ee7b19. --- subworkflows/local/binning.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 1788cbb9..65328eb0 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -77,6 +77,7 @@ workflow BINNING { [ meta_new, assembly, reads, depth ] } + .set { ch_maxbin2_input } ch_versions = ch_versions.mix(CONVERT_DEPTHS.out.versions.first()) } From 6d780eec86a5e32ecec3c5c86eed0bf14aaceba4 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 15:06:57 +0200 Subject: [PATCH 170/246] Revert "Merge branch 'dev' into dastool" This reverts commit 398e8794c0f695b871a0009eff2e35358a93d150, reversing changes made to 0e17759f7e01ff1e8449e640e250ad9be4de55dc. --- subworkflows/local/ancient_dna.nf | 3 +- subworkflows/local/binning.nf | 10 +++-- subworkflows/local/gtdbtk.nf | 6 ++- subworkflows/local/input_check.nf | 13 ++++--- workflows/mag.nf | 65 +++++++++++++++++++++---------- 5 files changed, 64 insertions(+), 33 deletions(-) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index fae88f82..c59c02e8 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -13,7 +13,8 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) FAIDX(input.map { item -> [ item[0], item[1] ] }) - freebayes_input = input.join(FAIDX.out.fai) // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] + input.join(FAIDX.out.fai) + .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], [], []] }, freebayes_input.map { item -> item[1] }, freebayes_input.map { item -> item[4] }, diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 65328eb0..52c27c33 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -44,13 +44,14 @@ workflow BINNING { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) - ch_metabat_depths = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth + METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth .map { meta, depths -> def meta_new = meta.clone() meta_new['binner'] = 'MetaBAT2' [ meta_new, depths ] } + .set { ch_metabat_depths } ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions.first()) @@ -70,7 +71,7 @@ workflow BINNING { // convert metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) - ch_maxbin2_input = CONVERT_DEPTHS.out.output + CONVERT_DEPTHS.out.output .map { meta, assembly, reads, depth -> def meta_new = meta.clone() meta_new['binner'] = 'MaxBin2' @@ -114,12 +115,13 @@ workflow BINNING { ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() } - ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() + SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) // mix all bins together - ch_final_bins_for_gunzip = ch_metabat2_results_transposed + ch_metabat2_results_transposed .mix( ch_maxbin2_results_transposed ) + .set { ch_final_bins_for_gunzip } GUNZIP_BINS ( ch_final_bins_for_gunzip ) GUNZIP_BINS.out.gunzip diff --git a/subworkflows/local/gtdbtk.nf b/subworkflows/local/gtdbtk.nf index fd971a33..2fe0e933 100644 --- a/subworkflows/local/gtdbtk.nf +++ b/subworkflows/local/gtdbtk.nf @@ -19,7 +19,7 @@ workflow GTDBTK { // Filter bins: classify only medium & high quality MAGs // Collect completness and contamination metrics from busco summary def bin_metrics = [:] - ch_busco_metrics = busco_summary + busco_summary .splitCsv(header: true, sep: '\t') .map { row -> def completeness = -1 @@ -36,9 +36,10 @@ workflow GTDBTK { if (duplicated != '') contamination = Double.parseDouble(duplicated) [row.'GenomeBin', completeness, contamination] } + .set { ch_busco_metrics } // Filter bins based on collected metrics: completeness, contamination - ch_filtered_bins = bins + bins .transpose() .map { meta, bin -> [bin.getName(), bin, meta]} .join(ch_busco_metrics, failOnDuplicate: true, failOnMismatch: true) @@ -49,6 +50,7 @@ workflow GTDBTK { discarded: (it[2] == -1 || it[2] < params.gtdbtk_min_completeness || it[3] == -1 || it[3] > params.gtdbtk_max_contamination) return [it[0], it[1]] } + .set { ch_filtered_bins } GTDBTK_DB_PREPARATION ( gtdb ) GTDBTK_CLASSIFY ( diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index 2e4af3de..348f4a16 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -10,7 +10,7 @@ workflow INPUT_CHECK { main: if(hasExtension(params.input, "csv")){ // extracts read files from samplesheet CSV and distribute into channels - ch_input_rows = Channel + Channel .from(file(params.input)) .splitCsv(header: true) .map { row -> @@ -30,9 +30,9 @@ workflow INPUT_CHECK { exit 1, "Input samplesheet contains row with ${row.size()} column(s). Expects 5." } } - + .set { ch_input_rows } // separate short and long reads - ch_raw_short_reads = ch_input_rows + ch_input_rows .map { id, group, sr1, sr2, lr -> def meta = [:] meta.id = id @@ -43,7 +43,8 @@ workflow INPUT_CHECK { else return [ meta, [ sr1, sr2 ] ] } - ch_raw_long_reads = ch_input_rows + .set { ch_raw_short_reads } + ch_input_rows .map { id, group, sr1, sr2, lr -> if (lr) { def meta = [:] @@ -52,8 +53,9 @@ workflow INPUT_CHECK { return [ meta, lr ] } } + .set { ch_raw_long_reads } } else { - ch_raw_short_reads = Channel + Channel .fromFilePairs(params.input, size: params.single_end ? 1 : 2) .ifEmpty { exit 1, "Cannot find any reads matching: ${params.input}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --single_end on the command line." } .map { row -> @@ -63,6 +65,7 @@ workflow INPUT_CHECK { meta.single_end = params.single_end return [ meta, row[1] ] } + .set { ch_raw_short_reads } ch_input_rows = Channel.empty() ch_raw_long_reads = Channel.empty() } diff --git a/workflows/mag.nf b/workflows/mag.nf index 3f37d4f9..dc5e3d3a 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -119,66 +119,78 @@ include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modu if ( params.host_genome ) { host_fasta = params.genomes[params.host_genome].fasta ?: false - ch_host_fasta = Channel + Channel .value(file( "${host_fasta}" )) + .set { ch_host_fasta } + host_bowtie2index = params.genomes[params.host_genome].bowtie2 ?: false - ch_host_bowtie2index = Channel + Channel .value(file( "${host_bowtie2index}/*" )) + .set { ch_host_bowtie2index } } else if ( params.host_fasta ) { - ch_host_fasta = Channel + Channel .value(file( "${params.host_fasta}" )) + .set { ch_host_fasta } } else { ch_host_fasta = Channel.empty() } if(params.busco_reference){ - ch_busco_db_file = Channel + Channel .value(file( "${params.busco_reference}" )) + .set { ch_busco_db_file } } else { ch_busco_db_file = Channel.empty() } if (params.busco_download_path) { - ch_busco_download_folder = Channel + Channel .value(file( "${params.busco_download_path}" )) + .set { ch_busco_download_folder } } else { ch_busco_download_folder = Channel.empty() } if(params.centrifuge_db){ - ch_centrifuge_db_file = Channel + Channel .value(file( "${params.centrifuge_db}" )) + .set { ch_centrifuge_db_file } } else { ch_centrifuge_db_file = Channel.empty() } if(params.kraken2_db){ - ch_kraken2_db_file = Channel + Channel .value(file( "${params.kraken2_db}" )) + .set { ch_kraken2_db_file } } else { ch_kraken2_db_file = Channel.empty() } if(params.cat_db){ - ch_cat_db_file = Channel + Channel .value(file( "${params.cat_db}" )) + .set { ch_cat_db_file } } else { ch_cat_db_file = Channel.empty() } if(!params.keep_phix) { - ch_phix_db_file = Channel + Channel .value(file( "${params.phix_reference}" )) + .set { ch_phix_db_file } } if (!params.keep_lambda) { - ch_nanolyse_db = Channel + Channel .value(file( "${params.lambda_reference}" )) + .set { ch_nanolyse_db } } gtdb = params.skip_busco ? false : params.gtdb if (gtdb) { - ch_gtdb = Channel + Channel .value(file( "${gtdb}" )) + .set { ch_gtdb } } else { ch_gtdb = Channel.empty() } @@ -358,12 +370,13 @@ workflow MAG { if (( params.centrifuge_db || params.kraken2_db ) && !params.skip_krona){ KRONA_DB () - ch_tax_classifications = CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) + CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) . map { classifier, meta, report -> def meta_new = meta.clone() meta_new.classifier = classifier [ meta_new, report ] } + .set { ch_tax_classifications } KRONA ( ch_tax_classifications, KRONA_DB.out.db.collect() @@ -381,7 +394,7 @@ workflow MAG { if (params.coassemble_group) { // short reads // group and set group as new id - ch_short_reads_grouped = ch_short_reads + ch_short_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -392,9 +405,10 @@ workflow MAG { if (!params.single_end) [ meta, reads.collect { it[0] }, reads.collect { it[1] } ] else [ meta, reads.collect { it }, [] ] } + .set { ch_short_reads_grouped } // long reads // group and set group as new id - ch_long_reads_grouped = ch_long_reads + ch_long_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -403,22 +417,25 @@ workflow MAG { meta.group = group [ meta, reads.collect { it } ] } + .set { ch_long_reads_grouped } } else { - ch_short_reads_grouped = ch_short_reads + ch_short_reads .map { meta, reads -> if (!params.single_end){ [ meta, [reads[0]], [reads[1]] ] } else [ meta, [reads], [] ] } + .set { ch_short_reads_grouped } } ch_assemblies = Channel.empty() if (!params.skip_megahit){ MEGAHIT ( ch_short_reads_grouped ) - ch_megahit_assemblies = MEGAHIT.out.assembly + MEGAHIT.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "MEGAHIT" [ meta_new, assembly ] } + .set { ch_megahit_assemblies } ch_assemblies = ch_assemblies.mix(ch_megahit_assemblies) ch_versions = ch_versions.mix(MEGAHIT.out.versions.first()) } @@ -442,36 +459,41 @@ workflow MAG { } } else { ch_short_reads_spades = ch_short_reads - ch_long_reads_spades = ch_long_reads + ch_long_reads .map { meta, reads -> [ meta, [reads] ] } + .set { ch_long_reads_spades } } if (!params.single_end && !params.skip_spades){ SPADES ( ch_short_reads_spades ) - ch_spades_assemblies = SPADES.out.assembly + SPADES.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdes" [ meta_new, assembly ] } + .set { ch_spades_assemblies } ch_assemblies = ch_assemblies.mix(ch_spades_assemblies) ch_versions = ch_versions.mix(SPADES.out.versions.first()) } if (!params.single_end && !params.skip_spadeshybrid){ - ch_short_reads_spades_tmp = ch_short_reads_spades + ch_short_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } - ch_reads_spadeshybrid = ch_long_reads_spades + .set {ch_short_reads_spades_tmp} + ch_long_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } .combine(ch_short_reads_spades_tmp, by: 0) .map { id, meta_long, long_reads, meta_short, short_reads -> [ meta_short, long_reads, short_reads ] } + .set { ch_reads_spadeshybrid } SPADESHYBRID ( ch_reads_spadeshybrid ) - ch_spadeshybrid_assemblies = SPADESHYBRID.out.assembly + SPADESHYBRID.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdesHybrid" [ meta_new, assembly ] } + .set { ch_spadeshybrid_assemblies } ch_assemblies = ch_assemblies.mix(ch_spadeshybrid_assemblies) ch_versions = ch_versions.mix(SPADESHYBRID.out.versions.first()) } @@ -665,6 +687,7 @@ workflow MAG { meta_new.id = bin.getBaseName() [ meta_new, bin ] } + .set { ch_bins_for_prokka } if (!params.skip_prokka){ PROKKA ( From a6f1c2a58a3b9da72d260240e29010d178cc9e82 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 4 May 2022 15:12:38 +0200 Subject: [PATCH 171/246] Account for when no binning refinement executed for bin_summary --- workflows/mag.nf | 77 ++++++++++++++++-------------------------------- 1 file changed, 26 insertions(+), 51 deletions(-) diff --git a/workflows/mag.nf b/workflows/mag.nf index dc5e3d3a..ee59e5d5 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -119,78 +119,66 @@ include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/modu if ( params.host_genome ) { host_fasta = params.genomes[params.host_genome].fasta ?: false - Channel + ch_host_fasta = Channel .value(file( "${host_fasta}" )) - .set { ch_host_fasta } - host_bowtie2index = params.genomes[params.host_genome].bowtie2 ?: false - Channel + ch_host_bowtie2index = Channel .value(file( "${host_bowtie2index}/*" )) - .set { ch_host_bowtie2index } } else if ( params.host_fasta ) { - Channel + ch_host_fasta = Channel .value(file( "${params.host_fasta}" )) - .set { ch_host_fasta } } else { ch_host_fasta = Channel.empty() } if(params.busco_reference){ - Channel + ch_busco_db_file = Channel .value(file( "${params.busco_reference}" )) - .set { ch_busco_db_file } } else { ch_busco_db_file = Channel.empty() } if (params.busco_download_path) { - Channel + ch_busco_download_folder = Channel .value(file( "${params.busco_download_path}" )) - .set { ch_busco_download_folder } } else { ch_busco_download_folder = Channel.empty() } if(params.centrifuge_db){ - Channel + ch_centrifuge_db_file = Channel .value(file( "${params.centrifuge_db}" )) - .set { ch_centrifuge_db_file } } else { ch_centrifuge_db_file = Channel.empty() } if(params.kraken2_db){ - Channel + ch_kraken2_db_file = Channel .value(file( "${params.kraken2_db}" )) - .set { ch_kraken2_db_file } } else { ch_kraken2_db_file = Channel.empty() } if(params.cat_db){ - Channel + ch_cat_db_file = Channel .value(file( "${params.cat_db}" )) - .set { ch_cat_db_file } } else { ch_cat_db_file = Channel.empty() } if(!params.keep_phix) { - Channel + ch_phix_db_file = Channel .value(file( "${params.phix_reference}" )) - .set { ch_phix_db_file } } if (!params.keep_lambda) { - Channel + ch_nanolyse_db = Channel .value(file( "${params.lambda_reference}" )) - .set { ch_nanolyse_db } } gtdb = params.skip_busco ? false : params.gtdb if (gtdb) { - Channel + ch_gtdb = Channel .value(file( "${gtdb}" )) - .set { ch_gtdb } } else { ch_gtdb = Channel.empty() } @@ -327,15 +315,13 @@ workflow MAG { } // join long and short reads by sample name - ch_short_reads + ch_short_reads_tmp = ch_short_reads .map { meta, sr -> [ meta.id, meta, sr ] } - .set {ch_short_reads_tmp} - ch_long_reads + ch_short_and_long_reads = ch_long_reads .map { meta, lr -> [ meta.id, meta, lr ] } .join(ch_short_reads_tmp, by: 0) .map { id, meta_lr, lr, meta_sr, sr -> [ meta_lr, lr, sr[0], sr[1] ] } // should not occur for single-end, since SPAdes (hybrid) does not support single-end - .set{ ch_short_and_long_reads } FILTLONG ( ch_short_and_long_reads @@ -370,13 +356,12 @@ workflow MAG { if (( params.centrifuge_db || params.kraken2_db ) && !params.skip_krona){ KRONA_DB () - CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) + ch_tax_classifications = CENTRIFUGE.out.results_for_krona.mix(KRAKEN2.out.results_for_krona) . map { classifier, meta, report -> def meta_new = meta.clone() meta_new.classifier = classifier [ meta_new, report ] } - .set { ch_tax_classifications } KRONA ( ch_tax_classifications, KRONA_DB.out.db.collect() @@ -394,7 +379,7 @@ workflow MAG { if (params.coassemble_group) { // short reads // group and set group as new id - ch_short_reads + ch_short_reads_grouped = ch_short_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -405,10 +390,9 @@ workflow MAG { if (!params.single_end) [ meta, reads.collect { it[0] }, reads.collect { it[1] } ] else [ meta, reads.collect { it }, [] ] } - .set { ch_short_reads_grouped } // long reads // group and set group as new id - ch_long_reads + ch_long_reads_grouped = ch_long_reads .map { meta, reads -> [ meta.group, meta, reads ] } .groupTuple(by: 0) .map { group, metas, reads -> @@ -417,25 +401,22 @@ workflow MAG { meta.group = group [ meta, reads.collect { it } ] } - .set { ch_long_reads_grouped } } else { - ch_short_reads + ch_short_reads_grouped = ch_short_reads .map { meta, reads -> if (!params.single_end){ [ meta, [reads[0]], [reads[1]] ] } else [ meta, [reads], [] ] } - .set { ch_short_reads_grouped } } ch_assemblies = Channel.empty() if (!params.skip_megahit){ MEGAHIT ( ch_short_reads_grouped ) - MEGAHIT.out.assembly + ch_megahit_assemblies = MEGAHIT.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "MEGAHIT" [ meta_new, assembly ] } - .set { ch_megahit_assemblies } ch_assemblies = ch_assemblies.mix(ch_megahit_assemblies) ch_versions = ch_versions.mix(MEGAHIT.out.versions.first()) } @@ -459,41 +440,36 @@ workflow MAG { } } else { ch_short_reads_spades = ch_short_reads - ch_long_reads + ch_long_reads_spades = ch_long_reads .map { meta, reads -> [ meta, [reads] ] } - .set { ch_long_reads_spades } } if (!params.single_end && !params.skip_spades){ SPADES ( ch_short_reads_spades ) - SPADES.out.assembly + ch_spades_assemblies = SPADES.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdes" [ meta_new, assembly ] } - .set { ch_spades_assemblies } ch_assemblies = ch_assemblies.mix(ch_spades_assemblies) ch_versions = ch_versions.mix(SPADES.out.versions.first()) } if (!params.single_end && !params.skip_spadeshybrid){ - ch_short_reads_spades + ch_short_reads_spades_tmp = ch_short_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } - .set {ch_short_reads_spades_tmp} - ch_long_reads_spades + ch_reads_spadeshybrid = ch_long_reads_spades .map { meta, reads -> [ meta.id, meta, reads ] } .combine(ch_short_reads_spades_tmp, by: 0) .map { id, meta_long, long_reads, meta_short, short_reads -> [ meta_short, long_reads, short_reads ] } - .set { ch_reads_spadeshybrid } SPADESHYBRID ( ch_reads_spadeshybrid ) - SPADESHYBRID.out.assembly + ch_spadeshybrid_assemblies = SPADESHYBRID.out.assembly .map { meta, assembly -> def meta_new = meta.clone() meta_new.assembler = "SPAdesHybrid" [ meta_new, assembly ] } - .set { ch_spadeshybrid_assemblies } ch_assemblies = ch_assemblies.mix(ch_spadeshybrid_assemblies) ch_versions = ch_versions.mix(SPADESHYBRID.out.versions.first()) } @@ -595,12 +571,12 @@ workflow MAG { ch_input_for_postbinning_bins = BINNING.out.bins.mix(BINNING_REFINEMENT.out.refined_bins) ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned,BINNING_REFINEMENT.out.refined_bins,BINNING_REFINEMENT.out.refined_unbins) ch_combinedepthtsvs_for_binsummary = BINNING.out.depths_summary.mix(BINNING_REFINEMENT.out.refined_depths_summary) - ch_input_for_binsummary = COMBINE_TSV ( ch_combinedepthtsvs_for_binsummary.collect() ).combined + ch_input_for_binsummary = COMBINE_TSV ( ch_combinedepthtsvs_for_binsummary.collect() ).combined } - } else { ch_input_for_postbinning_bins = BINNING.out.bins ch_input_for_postbinning_bins_unbins = BINNING.out.bins.mix(BINNING.out.unbinned) + ch_input_for_binsummary = BINNING.out.depths_summary } if (!params.skip_busco){ @@ -681,13 +657,12 @@ workflow MAG { /* * Prokka: Genome annotation */ - ch_input_for_postbinning_bins_unbins.transpose() + ch_bins_for_prokka = ch_input_for_postbinning_bins_unbins.transpose() .map { meta, bin -> def meta_new = meta.clone() meta_new.id = bin.getBaseName() [ meta_new, bin ] } - .set { ch_bins_for_prokka } if (!params.skip_prokka){ PROKKA ( From e59f544bbe7c8f1cf92e821d1d4b7d855517d58e Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 5 May 2022 13:55:16 +0200 Subject: [PATCH 172/246] Update documentation fix publishing for DASTOOL modules --- conf/modules.config | 6 +++--- docs/output.md | 12 ++++++++---- 2 files changed, 11 insertions(+), 7 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index b8bb2555..f77e8555 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -481,7 +481,7 @@ process { withName: DASTOOL_DASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinRefinement/" }, + path: { "${params.outdir}/GenomeBinningRefinement/" }, mode: params.publish_dir_mode, pattern: '*.{tsv,log,eval,seqlength}' ], @@ -493,7 +493,7 @@ process { withName: RENAME_PREDASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinRefinement/" }, + path: { "${params.outdir}/GenomeBinningRefinement/" }, mode: params.publish_dir_mode, pattern: '*.fa' ], @@ -504,7 +504,7 @@ process { withName: RENAME_POSTDASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinRefinement/" }, + path: { "${params.outdir}/GenomeBinningRefinement/" }, mode: params.publish_dir_mode, pattern: '*.fa' ], diff --git a/docs/output.md b/docs/output.md index 472b1d2a..bb24a016 100644 --- a/docs/output.md +++ b/docs/output.md @@ -329,7 +329,7 @@ Files in these two folders contain all contigs of an assembly. ### Bin sequencing depth -For each genome bin the median sequencing depth is computed based on the corresponding contig depths given in `GenomeBinning/[assembler]-[sample/group]-depth.txt.gz`. +For each genome bin the median sequencing depth is computed based on the corresponding contig depths.
Output files @@ -411,13 +411,17 @@ Besides the reference files or output files created by BUSCO, the following summ
Output files -- `BinRefinement/` +- `GenomeBinningRefinement/` + - `bin_depths_summary_refined.tsv`: Summary of bin sequencing depths for all samples against refined for refined bin contigs. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads + - `bin_summary.tsv`: Summary of bin and refined bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. This will be alternatively placed in `GenomeBinning/` when **no** binning refinement is performed - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig - - `[assembler]-[sample/group]_DASTool_bins/*`: Directory containing bins in FASTA format + - `[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format + - `[assembler]-DASToolUnbinned-[sample/group].*.fa`: Discarded contigs from bin refinement in fasta format + - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing refined bin abundances of the assembly across samples. Refined bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`.
@@ -500,7 +504,7 @@ Whole genome annotation is the process of identifying features of interest in a
Output files -- `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. +- `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. This will be alternatively placed in `GenomeBinningRefinement/` if `--refine_bins_dastool` binning refinement is performed.
From be51cb667a3a09ef7f7d957171707025813930c6 Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Thu, 5 May 2022 11:58:20 +0200 Subject: [PATCH 173/246] Apply suggestions from code review Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- conf/base.config | 4 ++-- conf/modules.config | 4 ++-- docs/usage.md | 2 -- 3 files changed, 4 insertions(+), 6 deletions(-) diff --git a/conf/base.config b/conf/base.config index 2abc49b5..d05a8912 100644 --- a/conf/base.config +++ b/conf/base.config @@ -161,12 +161,12 @@ process { } withName: MAXBIN2 { - // often fails when insufficient information, so we allow it to gracefully fail without filling the pipeline + // often fails when insufficient information, so we allow it to gracefully fail without failing the pipeline errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } } withName: DASTOOL_DASTOOL { - // if SCGs not found, bins cannot be assigned and DAS_tool will die + // if SCGs not found, bins cannot be assigned and DAS_tool will die with exit status 1 errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : task.exitStatus == 1 ? 'ignore' : 'finish' } } } diff --git a/conf/modules.config b/conf/modules.config index f77e8555..a907bff5 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -230,7 +230,7 @@ process { withName: 'MAG_DEPTHS_PLOT_REFINED|BIN_SUMMARY_REFINED' { publishDir = [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinningRefinement/" }, mode: params.publish_dir_mode, pattern: "*.{png,tsv}" ] @@ -239,7 +239,7 @@ process { withName: 'MAG_DEPTHS_SUMMARY_REFINED' { ext.prefix = "bin_depths_summary_refined" publishDir = [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinningRefinement/" }, mode: params.publish_dir_mode, pattern: "*.{tsv}" ] diff --git a/docs/usage.md b/docs/usage.md index 1198aaa0..0c6a4e72 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -299,5 +299,3 @@ DAS_Tool may not always be able to refine bins due to insufficient recovery of e In this case, DAS_Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the BinRefinement results directory for that sample. If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS_Tool publication](https://www.nature.com/articles/s41564-018-0171-1). - -Furthermore, only From 1e32d873f12493130cba36bc349c85de3b2b2fa1 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 9 May 2022 11:15:52 +0200 Subject: [PATCH 174/246] Replace .set operation with variable assignment again --- subworkflows/local/ancient_dna.nf | 3 +-- subworkflows/local/binning.nf | 26 +++++++++----------------- subworkflows/local/gtdbtk.nf | 6 ++---- subworkflows/local/input_check.nf | 12 ++++-------- 4 files changed, 16 insertions(+), 31 deletions(-) diff --git a/subworkflows/local/ancient_dna.nf b/subworkflows/local/ancient_dna.nf index c59c02e8..fae88f82 100644 --- a/subworkflows/local/ancient_dna.nf +++ b/subworkflows/local/ancient_dna.nf @@ -13,8 +13,7 @@ workflow ANCIENT_DNA_ASSEMLY_VALIDATION { PYDAMAGE_ANALYZE(input.map {item -> [item[0], item[2], item[3]]}) PYDAMAGE_FILTER(PYDAMAGE_ANALYZE.out.csv) FAIDX(input.map { item -> [ item[0], item[1] ] }) - input.join(FAIDX.out.fai) - .set { freebayes_input } // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] + freebayes_input = input.join(FAIDX.out.fai) // [val(meta), path(contigs), path(bam), path(bam_index), path(fai)] FREEBAYES (freebayes_input.map { item -> [item[0], item[2], item[3], [], [], []] }, freebayes_input.map { item -> item[1] }, freebayes_input.map { item -> item[4] }, diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 52c27c33..6dd5b117 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -44,14 +44,13 @@ workflow BINNING { METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS ( ch_summarizedepth_input ) - METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth + ch_metabat_depths = METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth .map { meta, depths -> def meta_new = meta.clone() meta_new['binner'] = 'MetaBAT2' [ meta_new, depths ] } - .set { ch_metabat_depths } ch_versions = ch_versions.mix(METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.versions.first()) @@ -71,14 +70,13 @@ workflow BINNING { // convert metabat2 depth files to maxbin2 if ( !params.skip_maxbin2 ) { CONVERT_DEPTHS ( ch_metabat2_input ) - CONVERT_DEPTHS.out.output + ch_maxbin2_input = CONVERT_DEPTHS.out.output .map { meta, assembly, reads, depth -> def meta_new = meta.clone() meta_new['binner'] = 'MaxBin2' [ meta_new, assembly, reads, depth ] } - .set { ch_maxbin2_input } ch_versions = ch_versions.mix(CONVERT_DEPTHS.out.versions.first()) } @@ -115,28 +113,25 @@ workflow BINNING { ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() } - SPLIT_FASTA.out.unbinned.transpose().set { ch_split_fasta_results_transposed } + ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) // mix all bins together - ch_metabat2_results_transposed + ch_final_bins_for_gunzip = ch_metabat2_results_transposed .mix( ch_maxbin2_results_transposed ) - .set { ch_final_bins_for_gunzip } GUNZIP_BINS ( ch_final_bins_for_gunzip ) - GUNZIP_BINS.out.gunzip - .set{ ch_binning_results_gunzipped } + ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions.first()) GUNZIP_UNBINS ( ch_split_fasta_results_transposed ) - GUNZIP_UNBINS.out.gunzip - .set{ ch_splitfasta_results_gunzipped } + ch_splitfasta_results_gunzipped = GUNZIP_UNBINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions.first()) // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta for grouping to mix back with original depth // files, as required for MAG_DEPTHS - ch_binning_results_gunzipped + ch_depth_input = ch_binning_results_gunzipped .mix(ch_splitfasta_results_gunzipped ) .map { meta, results -> def meta_new = meta.clone() @@ -145,7 +140,6 @@ workflow BINNING { } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) - .set { ch_depth_input } MAG_DEPTHS ( ch_depth_input ) ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) @@ -171,13 +165,11 @@ workflow BINNING { ch_versions = ch_versions.mix( MAG_DEPTHS_SUMMARY.out.versions ) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped + ch_binning_results_gunzipped_final = ch_binning_results_gunzipped .groupTuple(by: 0) - .set{ ch_binning_results_gunzipped_final } - METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) + ch_binning_results_gzipped_final = METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) .groupTuple(by: 0) - .set{ ch_binning_results_gzipped_final } SPLIT_FASTA.out.unbinned diff --git a/subworkflows/local/gtdbtk.nf b/subworkflows/local/gtdbtk.nf index 2fe0e933..fd971a33 100644 --- a/subworkflows/local/gtdbtk.nf +++ b/subworkflows/local/gtdbtk.nf @@ -19,7 +19,7 @@ workflow GTDBTK { // Filter bins: classify only medium & high quality MAGs // Collect completness and contamination metrics from busco summary def bin_metrics = [:] - busco_summary + ch_busco_metrics = busco_summary .splitCsv(header: true, sep: '\t') .map { row -> def completeness = -1 @@ -36,10 +36,9 @@ workflow GTDBTK { if (duplicated != '') contamination = Double.parseDouble(duplicated) [row.'GenomeBin', completeness, contamination] } - .set { ch_busco_metrics } // Filter bins based on collected metrics: completeness, contamination - bins + ch_filtered_bins = bins .transpose() .map { meta, bin -> [bin.getName(), bin, meta]} .join(ch_busco_metrics, failOnDuplicate: true, failOnMismatch: true) @@ -50,7 +49,6 @@ workflow GTDBTK { discarded: (it[2] == -1 || it[2] < params.gtdbtk_min_completeness || it[3] == -1 || it[3] > params.gtdbtk_max_contamination) return [it[0], it[1]] } - .set { ch_filtered_bins } GTDBTK_DB_PREPARATION ( gtdb ) GTDBTK_CLASSIFY ( diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index 348f4a16..53a5103c 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -10,7 +10,7 @@ workflow INPUT_CHECK { main: if(hasExtension(params.input, "csv")){ // extracts read files from samplesheet CSV and distribute into channels - Channel + ch_input_rows = Channel .from(file(params.input)) .splitCsv(header: true) .map { row -> @@ -30,9 +30,8 @@ workflow INPUT_CHECK { exit 1, "Input samplesheet contains row with ${row.size()} column(s). Expects 5." } } - .set { ch_input_rows } // separate short and long reads - ch_input_rows + ch_raw_short_reads = ch_input_rows .map { id, group, sr1, sr2, lr -> def meta = [:] meta.id = id @@ -43,8 +42,7 @@ workflow INPUT_CHECK { else return [ meta, [ sr1, sr2 ] ] } - .set { ch_raw_short_reads } - ch_input_rows + ch_raw_long_reads = ch_input_rows .map { id, group, sr1, sr2, lr -> if (lr) { def meta = [:] @@ -53,9 +51,8 @@ workflow INPUT_CHECK { return [ meta, lr ] } } - .set { ch_raw_long_reads } } else { - Channel + ch_raw_short_reads = Channel .fromFilePairs(params.input, size: params.single_end ? 1 : 2) .ifEmpty { exit 1, "Cannot find any reads matching: ${params.input}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --single_end on the command line." } .map { row -> @@ -65,7 +62,6 @@ workflow INPUT_CHECK { meta.single_end = params.single_end return [ meta, row[1] ] } - .set { ch_raw_short_reads } ch_input_rows = Channel.empty() ch_raw_long_reads = Channel.empty() } From eb728e1b0b48f4a32a332dfdbc1bd4caf95bfb45 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 9 May 2022 13:37:20 +0200 Subject: [PATCH 175/246] Remove unnecessary binner info in meta after RENAME_PREDASTOOL --- subworkflows/local/binning_refinement.nf | 12 ++---------- 1 file changed, 2 insertions(+), 10 deletions(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 75163538..bd5d89af 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -37,18 +37,10 @@ workflow BINNING_REFINEMENT { [ meta_new, assembly ] } - // Rename for consistency for downstream steps (BUSCO etc.) ch_bins_for_fastatocontig2bin = RENAME_PREDASTOOL(bins).renamed_bins - .map { - meta, bins -> - def meta_new = meta.clone() - meta_new.binner = meta.binner + 'Refined' - - [ meta_new, bins ] - } .branch { - metabat2: it[0]['binner'] == 'MetaBAT2Refined' - maxbin2: it[0]['binner'] == 'MaxBin2Refined' + metabat2: it[0]['binner'] == 'MetaBAT2' + maxbin2: it[0]['binner'] == 'MaxBin2' } // Generate DASTool auxilary files From 22cf70ed8b39b621e109e0dc04cd04f95cefaae3 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 9 May 2022 15:38:54 +0200 Subject: [PATCH 176/246] Collapse binning and refinement output directories: note this will result in overwriting of various summary files (heatmap, depth files, possibly QUAST) --- conf/modules.config | 27 ++++++++++--------------- docs/output.md | 48 ++++++++++++++++++++++----------------------- 2 files changed, 33 insertions(+), 42 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index a907bff5..44591841 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -222,7 +222,7 @@ process { // GenomeBinning directory withName: 'BIN_SUMMARY' { publishDir = [ - path: params.refine_bins_dastool ? { "${params.outdir}/GenomeBinningRefinement" } : { "${params.outdir}/GenomeBinning" }, + path: { "${params.outdir}/GenomeBinning" }, mode: params.publish_dir_mode, pattern: "*.{png,tsv}" ] @@ -230,7 +230,7 @@ process { withName: 'MAG_DEPTHS_PLOT_REFINED|BIN_SUMMARY_REFINED' { publishDir = [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinning/" }, mode: params.publish_dir_mode, pattern: "*.{png,tsv}" ] @@ -239,7 +239,7 @@ process { withName: 'MAG_DEPTHS_SUMMARY_REFINED' { ext.prefix = "bin_depths_summary_refined" publishDir = [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinning/" }, mode: params.publish_dir_mode, pattern: "*.{tsv}" ] @@ -255,12 +255,13 @@ process { withName: 'BUSCO|BUSCO_PLOT' { publishDir = [ - path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] } + withName: BUSCO_SAVE_DOWNLOAD { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, @@ -270,14 +271,6 @@ process { ] } - withName: 'BUSCO_SUMMARY|QUAST_BINS|QUAST_BINS_SUMMARY' { - publishDir = [ - path: { "${params.outdir}/GenomeBinning/QC" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] - } - withName: CAT_DB_GENERATE { publishDir = [ path: { "${params.outdir}/Taxonomy/CAT" }, @@ -481,7 +474,7 @@ process { withName: DASTOOL_DASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinning/DASTool" }, mode: params.publish_dir_mode, pattern: '*.{tsv,log,eval,seqlength}' ], @@ -493,7 +486,7 @@ process { withName: RENAME_PREDASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinning/DASTool/bins" }, mode: params.publish_dir_mode, pattern: '*.fa' ], @@ -504,7 +497,7 @@ process { withName: RENAME_POSTDASTOOL { publishDir = [ [ - path: { "${params.outdir}/GenomeBinningRefinement/" }, + path: { "${params.outdir}/GenomeBinning/DASTool/unbinned" }, mode: params.publish_dir_mode, pattern: '*.fa' ], diff --git a/docs/output.md b/docs/output.md index bb24a016..facda0e1 100644 --- a/docs/output.md +++ b/docs/output.md @@ -252,7 +252,7 @@ Protein-coding genes are predicted for each assembly.
-## Binning +## Binning and Binning Refinement ### Contig sequencing depth @@ -327,6 +327,25 @@ All the files in this folder contain small and/or unbinned contigs that are not Files in these two folders contain all contigs of an assembly. +### DAS Tool + +[DAS Tool](https://github.com/cmks/DAS_Tool) is an automated binning refinement method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. nf-core/mag uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins. + +
+Output files + +- `GenomeBinning/DASTool/` + - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets + - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets + - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners + - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information + - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig + - `bins/[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format + - `bins/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format + - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing refined bin abundances of the assembly across samples. Refined bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. + +
+ ### Bin sequencing depth For each genome bin the median sequencing depth is computed based on the corresponding contig depths. @@ -336,6 +355,7 @@ For each genome bin the median sequencing depth is computed based on the corresp - `GenomeBinning/depths/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. + - `bin_depths_summary_refined.tsv`: Summary of bin sequencing depths for all samples against refined bin contigs, if refinement performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`.
@@ -367,7 +387,7 @@ For each genome bin the median sequencing depth is computed based on the corresp ### QC for metagenome assembled genomes with BUSCO -[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and high quality contigs obtained by MetaBAT2. By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated. +[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and refined bins and high quality contigs obtained by MetaBAT2. By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated.
Output files @@ -403,28 +423,6 @@ Besides the reference files or output files created by BUSCO, the following summ - `busco_summary.tsv`: A summary table of the BUSCO results, with % of marker genes found. If run in automated lineage selection mode, both the results for the selected domain and for the selected more specific lineage will be given, if available.
- -### Bin Refinement - -[DAS Tool](https://github.com/cmks/DAS_Tool) is an automated method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. nf-core/mag uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins. - -
-Output files - -- `GenomeBinningRefinement/` - - `bin_depths_summary_refined.tsv`: Summary of bin sequencing depths for all samples against refined for refined bin contigs. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads - - `bin_summary.tsv`: Summary of bin and refined bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. This will be alternatively placed in `GenomeBinning/` when **no** binning refinement is performed - - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets - - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets - - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners - - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information - - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig - - `[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format - - `[assembler]-DASToolUnbinned-[sample/group].*.fa`: Discarded contigs from bin refinement in fasta format - - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing refined bin abundances of the assembly across samples. Refined bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. - -
- ## Taxonomic classification of binned genomes ### CAT @@ -504,7 +502,7 @@ Whole genome annotation is the process of identifying features of interest in a
Output files -- `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. This will be alternatively placed in `GenomeBinningRefinement/` if `--refine_bins_dastool` binning refinement is performed. +- `GenomeBinning/bin_summary.tsv`: Summary of bin sequencing depths together with BUSCO, QUAST and GTDB-Tk results, if at least one of the later was generated. This will also include refined bins if `--refine_bins_dastool` binning refinement is performed.
From cf8bc402453a3ea9a5e4e584a34a41c97b2b53be Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Mon, 16 May 2022 10:18:36 +0000 Subject: [PATCH 177/246] Template update for nf-core/tools version 2.4 --- .github/workflows/awsfulltest.yml | 3 - .github/workflows/awstest.yml | 3 - .github/workflows/branch.yml | 3 +- .github/workflows/ci.yml | 2 - .github/workflows/fix-linting.yml | 55 ++++++++++ .github/workflows/linting.yml | 4 +- .github/workflows/linting_comment.yml | 1 - .prettierignore | 9 ++ README.md | 25 ++--- assets/email_template.html | 142 ++++++++------------------ bin/check_samplesheet.py | 16 ++- nextflow.config | 2 +- 12 files changed, 135 insertions(+), 130 deletions(-) create mode 100644 .github/workflows/fix-linting.yml create mode 100644 .prettierignore diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 414181d9..e7365baf 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -28,6 +28,3 @@ jobs: "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-${{ github.sha }}" } profiles: test_full,aws_tower - nextflow_config: | - process.errorStrategy = 'retry' - process.maxRetries = 3 diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index c277a5cb..e5f69c5d 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -23,6 +23,3 @@ jobs: "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/mag/results-test-${{ github.sha }}" } profiles: test,aws_tower - nextflow_config: | - process.errorStrategy = 'retry' - process.maxRetries = 3 diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 55f14919..e08454a2 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,7 +13,7 @@ jobs: - name: Check PRs if: github.repository == 'nf-core/mag' run: | - "{ [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/mag ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]" + { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/mag ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] # If the above check failed, post a comment on the PR explaining the failure # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets @@ -42,4 +42,3 @@ jobs: Thanks again for your contribution! repo-token: ${{ secrets.GITHUB_TOKEN }} allow-repeats: false -# diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index f5885cbc..2d7a9053 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -48,5 +48,3 @@ jobs: # Remember that you can parallelise this by using strategy.matrix run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results - -# diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml new file mode 100644 index 00000000..eca5edd4 --- /dev/null +++ b/.github/workflows/fix-linting.yml @@ -0,0 +1,55 @@ +name: Fix linting from a comment +on: + issue_comment: + types: [created] + +jobs: + deploy: + # Only run if comment is on a PR with the main repo, and if it contains the magic keywords + if: > + contains(github.event.comment.html_url, '/pull/') && + contains(github.event.comment.body, '@nf-core-bot fix linting') && + github.repository == 'nf-core/mag' + runs-on: ubuntu-latest + steps: + # Use the @nf-core-bot token to check out so we can push later + - uses: actions/checkout@v3 + with: + token: ${{ secrets.nf_core_bot_auth_token }} + + # Action runs on the issue comment, so we don't get the PR by default + # Use the gh cli to check out the PR + - name: Checkout Pull Request + run: gh pr checkout ${{ github.event.issue.number }} + env: + GITHUB_TOKEN: ${{ secrets.nf_core_bot_auth_token }} + + - uses: actions/setup-node@v2 + + - name: Install Prettier + run: npm install -g prettier @prettier/plugin-php + + # Check that we actually need to fix something + - name: Run 'prettier --check' + id: prettier_status + run: | + if prettier --check ${GITHUB_WORKSPACE}; then + echo "::set-output name=result::pass" + else + echo "::set-output name=result::fail" + fi + + - name: Run 'prettier --write' + if: steps.prettier_status.outputs.result == 'fail' + run: prettier --write ${GITHUB_WORKSPACE} + + - name: Commit & push changes + if: steps.prettier_status.outputs.result == 'fail' + run: | + git config user.email "core@nf-co.re" + git config user.name "nf-core-bot" + git config push.default upstream + git add . + git status + git commit -m "[automated] Fix linting with Prettier" + git push diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index e9cf5de3..77358dee 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -48,7 +48,7 @@ jobs: wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ - - uses: actions/setup-python@v1 + - uses: actions/setup-python@v3 with: python-version: "3.6" architecture: "x64" @@ -78,5 +78,3 @@ jobs: lint_log.txt lint_results.md PR_number.txt - -# diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml index 91c487a1..04758f61 100644 --- a/.github/workflows/linting_comment.yml +++ b/.github/workflows/linting_comment.yml @@ -26,4 +26,3 @@ jobs: GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} number: ${{ steps.pr_number.outputs.pr_number }} path: linting-logs/lint_results.md -# diff --git a/.prettierignore b/.prettierignore new file mode 100644 index 00000000..d0e7ae58 --- /dev/null +++ b/.prettierignore @@ -0,0 +1,9 @@ +email_template.html +.nextflow* +work/ +data/ +results/ +.DS_Store +testing/ +testing* +*.pyc diff --git a/README.md b/README.md index 8819a575..bf57fbc3 100644 --- a/README.md +++ b/README.md @@ -1,18 +1,19 @@ -# ![nf-core/mag](docs/images/nf-core/mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core/mag_logo_dark.png#gh-dark-mode-only) +# ![nf-core/mag](docs/images/nf-core-mag_logo_light.png#gh-light-mode-only) ![nf-core/mag](docs/images/nf-core-mag_logo_dark.png#gh-dark-mode-only) [![GitHub Actions CI Status](https://github.com/nf-core/mag/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/mag/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/mag/actions?query=workflow%3A%22nf-core+linting%22) -[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/mag/results) -[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) - -[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.10.3-23aa62.svg?labelColor=000000)](https://www.nextflow.io/) -[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) -[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) -[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) - -[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23mag-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/mag) -[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core) -[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) +[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?logo=Amazon%20AWS)](https://nf-co.re/mag/results) +[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8)](https://doi.org/10.5281/zenodo.XXXXXXX) + +[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A521.10.3-23aa62.svg)](https://www.nextflow.io/) +[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?logo=anaconda)](https://docs.conda.io/en/latest/) +[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?logo=docker)](https://www.docker.com/) +[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg)](https://sylabs.io/docs/) +[![Launch on Nextflow Tower](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Nextflow%20Tower-%234256e7)](https://tower.nf/launch?pipeline=https://github.com/nf-core/mag) + +[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23mag-4A154B?logo=slack)](https://nfcore.slack.com/channels/mag) +[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?logo=twitter)](https://twitter.com/nf_core) +[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?logo=youtube)](https://www.youtube.com/c/nf-core) ## Introduction diff --git a/assets/email_template.html b/assets/email_template.html index 1563166a..13cdbf26 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -1,111 +1,53 @@ - - - - + + + + - - - nf-core/mag Pipeline Report - - -
- + + nf-core/mag Pipeline Report + + +
-

nf-core/mag v${version}

-

Run Name: $runName

+ - <% if (!success){ out << """ -
-

nf-core/mag execution completed unsuccessfully!

+

nf-core/mag v${version}

+

Run Name: $runName

+ +<% if (!success){ + out << """ +
+

nf-core/mag execution completed unsuccessfully!

The exit status of the task that caused the workflow execution to fail was: $exitStatus.

The full error message was:

- 
${errorReport}
-
- """ } else { out << """ -
+ 
${errorReport}
+
+ """ +} else { + out << """ +
nf-core/mag execution completed successfully! -
- """ } %> +
+ """ +} +%> -

The workflow was completed at $dateComplete (duration: $duration)

-

The command used to launch the workflow was as follows:

- 
-$commandLine
+

The workflow was completed at $dateComplete (duration: $duration)

+

The command used to launch the workflow was as follows:

+
$commandLine
-

Pipeline Configuration:

- - - <% out << summary.collect{ k,v -> " - - - - - " }.join("\n") %> - -
- $k - - 
$v
-
+

Pipeline Configuration:

+ + + <% out << summary.collect{ k,v -> "" }.join("\n") %> + +
$k
$v
-

nf-core/mag

-

https://github.com/nf-core/mag

-
- +

nf-core/mag

+

https://github.com/nf-core/mag

+ +
+ + diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index 5473b624..3652c63c 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -98,7 +98,7 @@ def _validate_pair(self, row): if row[self._first_col] and row[self._second_col]: row[self._single_col] = False assert ( - Path(row[self._first_col]).suffixes == Path(row[self._second_col]).suffixes + Path(row[self._first_col]).suffixes[-2:] == Path(row[self._second_col]).suffixes[-2:] ), "FASTQ pairs must have the same file extensions." else: row[self._single_col] = True @@ -129,6 +129,16 @@ def validate_unique_samples(self): row[self._sample_col] = f"{sample}_T{seen[sample]}" +def read_head(handle, num_lines=10): + """Read the specified number of lines from the current position in the file.""" + lines = [] + for idx, line in enumerate(handle): + if idx == num_lines: + break + lines.append(line) + return "".join(lines) + + def sniff_format(handle): """ Detect the tabular format. @@ -144,13 +154,13 @@ def sniff_format(handle): https://docs.python.org/3/glossary.html#term-text-file """ - peek = handle.read(2048) + peek = read_head(handle) + handle.seek(0) sniffer = csv.Sniffer() if not sniffer.has_header(peek): logger.critical(f"The given sample sheet does not appear to contain a header.") sys.exit(1) dialect = sniffer.sniff(peek) - handle.seek(0) return dialect diff --git a/nextflow.config b/nextflow.config index 50dbaf7e..ffb73b89 100644 --- a/nextflow.config +++ b/nextflow.config @@ -159,7 +159,7 @@ trace { } dag { enabled = true - file = "${params.tracedir}/pipeline_dag_${trace_timestamp}.svg" + file = "${params.tracedir}/pipeline_dag_${trace_timestamp}.html" } manifest { From 1a872df3982f76af0aa63bad353a4f2a39b0bccd Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 15:54:26 +0200 Subject: [PATCH 178/246] Add module publishing dirs for BUSCO_SUMMARY|QUAST_BINS|QUAST_BINS_SUMMARY again --- conf/modules.config | 9 ++++++++- 1 file changed, 8 insertions(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 44591841..c962115c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -261,7 +261,6 @@ process { ] } - withName: BUSCO_SAVE_DOWNLOAD { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, @@ -271,6 +270,14 @@ process { ] } + withName: 'BUSCO_SUMMARY|QUAST_BINS|QUAST_BINS_SUMMARY' { + publishDir = [ + path: { "${params.outdir}/GenomeBinning/QC" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + withName: CAT_DB_GENERATE { publishDir = [ path: { "${params.outdir}/Taxonomy/CAT" }, From e9e81ce06a5186b47a800e8087ecd04473396463 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 16:32:55 +0200 Subject: [PATCH 179/246] Clean up modules.config --- conf/modules.config | 29 +++++------------------------ 1 file changed, 5 insertions(+), 24 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index c962115c..df449e2a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -201,7 +201,7 @@ process { ] } - withName: 'MAG_DEPTHS_PLOT|MAG_DEPTHS_SUMMARY' { + withName: 'MAG_DEPTHS_PLOT|MAG_DEPTHS_SUMMARY|MAG_DEPTHS_PLOT_REFINED' { publishDir = [ path: { "${params.outdir}/GenomeBinning/depths" }, mode: params.publish_dir_mode, @@ -209,17 +209,15 @@ process { ] } - withName: 'BIN_SUMMARY' { + withName: 'MAG_DEPTHS_SUMMARY_REFINED' { + ext.prefix = "bin_depths_summary_refined" publishDir = [ - path: { "${params.outdir}/GenomeBinning" }, + path: { "${params.outdir}/GenomeBinning/depths" }, mode: params.publish_dir_mode, - pattern: "*.{png,tsv}" + pattern: "*.{tsv}" ] } - // Always push the publishDir for this step to the further downstream as - // possible, as it doesn't make sense to included refined bins in just the - // GenomeBinning directory withName: 'BIN_SUMMARY' { publishDir = [ path: { "${params.outdir}/GenomeBinning" }, @@ -228,23 +226,6 @@ process { ] } - withName: 'MAG_DEPTHS_PLOT_REFINED|BIN_SUMMARY_REFINED' { - publishDir = [ - path: { "${params.outdir}/GenomeBinning/" }, - mode: params.publish_dir_mode, - pattern: "*.{png,tsv}" - ] - } - - withName: 'MAG_DEPTHS_SUMMARY_REFINED' { - ext.prefix = "bin_depths_summary_refined" - publishDir = [ - path: { "${params.outdir}/GenomeBinning/" }, - mode: params.publish_dir_mode, - pattern: "*.{tsv}" - ] - } - withName: BUSCO_DB_PREPARATION { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, From d1b7d1245bc22391466c79605c40b96866f2339f Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 16:34:51 +0200 Subject: [PATCH 180/246] Fix how-to-run comment in test_binrefinement.config --- conf/test_binrefinement.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_binrefinement.config b/conf/test_binrefinement.config index 1931dd11..ddf44ceb 100644 --- a/conf/test_binrefinement.config +++ b/conf/test_binrefinement.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test, --outdir + nextflow run nf-core/mag -profile test_binrefinement, --outdir ---------------------------------------------------------------------------------------- */ From b29c2b71e6e0d556ddd1f933fff0c54d32a88446 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 16:45:42 +0200 Subject: [PATCH 181/246] Simplify code snippet to remove binner meta info --- subworkflows/local/binning_refinement.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index bd5d89af..5d89f2bb 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -96,7 +96,8 @@ workflow BINNING_REFINEMENT { .mix( RENAME_POSTDASTOOL.out.refined_unbins ) .map { meta, refinedbins -> - def meta_new = [ 'id': meta.id, 'group': meta.group, 'single_end': meta.single_end, 'assembler': meta.assembler ] + def meta_new = meta.clone() + meta_new.remove('binner') [ meta_new, refinedbins ] } .join( depths, by: 0 ) From b44ff202f550cf701a4c5ec81866ce7cc2277bfb Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 16:46:55 +0200 Subject: [PATCH 182/246] Adjust binning_refinement.nf for separate plotting for binners --- subworkflows/local/binning_refinement.nf | 8 +++++++- 1 file changed, 7 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 5d89f2bb..a0e666c3 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -109,9 +109,15 @@ workflow BINNING_REFINEMENT { ch_sample_groups = reads .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } + // Transpose and add 'binner' meta information again for plotting // filter MAG depth files: use only those for plotting that contain depths for > 2 samples ch_mag_depths_plot_refined = MAG_DEPTHS_REFINED.out.depths - .map { meta, depth_file -> if (getColNo(depth_file) > 2) [meta, depth_file] } + .transpose() + .map { meta, depth_file -> + def meta_new = meta.clone() + meta_new['binner'] = depth_file.name.split("-")[1] + if (getColNo(depth_file) > 2) [meta_new, depth_file] + } MAG_DEPTHS_PLOT_REFINED ( ch_mag_depths_plot_refined, ch_sample_groups.collect() ) MAG_DEPTHS_SUMMARY_REFINED ( MAG_DEPTHS_REFINED.out.depths.map{it[1]}.collect() ) From 0cdb5ddeace604e99c2ea6bfca910e5d430b4e53 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 17:14:18 +0200 Subject: [PATCH 183/246] Adjust output description of DAS Tool to new structure --- docs/output.md | 1 - 1 file changed, 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index facda0e1..45fc6abb 100644 --- a/docs/output.md +++ b/docs/output.md @@ -342,7 +342,6 @@ Files in these two folders contain all contigs of an assembly. - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig - `bins/[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format - `bins/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format - - `[assembler]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing refined bin abundances of the assembly across samples. Refined bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`.
From 02a582a1ae9f350baebbec4bb4a7e8ed5c95431d Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 17:17:48 +0200 Subject: [PATCH 184/246] Update BUSCO description in output.md for new binning and refinment methods --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 45fc6abb..b8ac15fc 100644 --- a/docs/output.md +++ b/docs/output.md @@ -386,7 +386,7 @@ For each genome bin the median sequencing depth is computed based on the corresp ### QC for metagenome assembled genomes with BUSCO -[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and refined bins and high quality contigs obtained by MetaBAT2. By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated. +[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and high quality contigs obtained by the applied binning and binning refinement methods. By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated.
Output files From 1b7cb58c3d2295fa959bd4dca6b333ab7918eeae Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 21:28:10 +0200 Subject: [PATCH 185/246] Fix undefined channels and restructure code --- subworkflows/local/binning.nf | 30 +++++++++++++----------------- 1 file changed, 13 insertions(+), 17 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 7c57c7ae..06f498b9 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -79,14 +79,22 @@ workflow BINNING { } } + // main bins for decompressing for MAG_DEPTHS + ch_final_bins_for_gunzip = Channel.empty() + // final gzipped bins + ch_binning_results_gzipped_final = Channel.empty() // run binning if ( !params.skip_metabat2 ) { METABAT2_METABAT2 ( ch_metabat2_input ) + // before decompressing first have to separate and re-group due to limitation of GUNZIP module + ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( METABAT2_METABAT2.out.fasta.transpose() ) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( METABAT2_METABAT2.out.fasta ) ch_versions = ch_versions.mix(METABAT2_METABAT2.out.versions) } - if ( !params.skip_maxbin2 ) { MAXBIN2 ( ch_maxbin2_input ) + ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( MAXBIN2.out.binned_fastas.transpose() ) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( MAXBIN2.out.binned_fastas ) ch_versions = ch_versions.mix(MAXBIN2.out.versions) } @@ -100,20 +108,11 @@ workflow BINNING { } SPLIT_FASTA ( ch_input_splitfasta ) - - // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for - // MAG_DEPTHS, first have to separate and re-group due to limitation of - // GUNZIP module - ch_metabat2_results_transposed = METABAT2_METABAT2.out.fasta.transpose() - ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() - + // large unbinned contigs from SPLIT_FASTA for decompressing for MAG_DEPTHS, + // first have to separate and re-group due to limitation of GUNZIP module ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) - // mix all bins together - ch_final_bins_for_gunzip = ch_metabat2_results_transposed - .mix( ch_maxbin2_results_transposed ) - GUNZIP_BINS ( ch_final_bins_for_gunzip ) ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions) @@ -159,11 +158,8 @@ workflow BINNING { ch_versions = ch_versions.mix(MAG_DEPTHS_SUMMARY.out.versions) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped_final = ch_binning_results_gunzipped - .groupTuple(by: 0) - - ch_binning_results_gzipped_final = METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) - .groupTuple(by: 0) + ch_binning_results_gunzipped_final = ch_binning_results_gunzipped.groupTuple(by: 0) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.groupTuple(by: 0) SPLIT_FASTA.out.unbinned From 021d695f64b2d452bb360e55c5263589f7b59cdf Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 10:16:57 +0200 Subject: [PATCH 186/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index be27e2d5..0580ab0d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,6 +17,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289) - Update to nf-core 2.3.2 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint - [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths` for `[assembler]-[sample/group]-depth.txt.gz`, `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` +- [#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` ### `Fixed` From f8f04c4dc1d13e9dd5dad82b380dc5e4513733c0 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 13:28:18 +0200 Subject: [PATCH 187/246] Apply suggestions from code review --- CHANGELOG.md | 2 +- CITATIONS.md | 2 +- README.md | 2 +- docs/usage.md | 6 +++--- modules/local/combine_tsv.nf | 4 ++-- modules/local/mag_depths_summary.nf | 2 +- modules/local/rename_postdastool.nf | 2 +- modules/local/rename_predastool.nf | 2 +- nextflow_schema.json | 8 ++++---- workflows/mag.nf | 2 +- 10 files changed, 16 insertions(+), 16 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 98be240c..a3eb1a8d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,7 +11,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool - [#284](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer -- [#284](https://github.com/nf-core/mag/pull/291) - Add DAS_Tool for bin refinement +- [#284](https://github.com/nf-core/mag/pull/291) - Add DAS Tool for bin refinement ### `Changed` diff --git a/CITATIONS.md b/CITATIONS.md index 8d9ac2ac..33337945 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -34,7 +34,7 @@ > Kim, D., Song, L., Breitwieser, F. P., & Salzberg, S. L. (2016). Centrifuge: rapid and sensitive classification of metagenomic sequences. Genome research, 26(12), 1721-1729. doi: 10.1101/gr.210641.116. -- [DAS_Tool](https://doi.org/10.1038/s41564-018-0171-1) +- [DAS Tool](https://doi.org/10.1038/s41564-018-0171-1) > Sieber, C. M. K., et al. 2018. "Recovery of Genomes from Metagenomes via a Dereplication, Aggregation and Scoring Strategy." Nature Microbiology 3 (7): 836-43. doi: 10.1038/s41564-018-0171-1 diff --git a/README.md b/README.md index 79ed234f..1d68e974 100644 --- a/README.md +++ b/README.md @@ -38,7 +38,7 @@ The pipeline then: - (optionally) performs ancient DNA assembly validation using [PyDamage](https://github.com/maxibor/pydamage) and contig consensus sequence recalling with [Freebayes](https://github.com/freebayes/freebayes) and [BCFtools](http://samtools.github.io/bcftools/bcftools.html) - predicts protein-coding genes for the assemblies using [Prodigal](https://github.com/hyattpd/Prodigal) - performs metagenome binning using [MetaBAT2](https://bitbucket.org/berkeleylab/metabat/src/master/) and/or with [MaxBin2](https://sourceforge.net/projects/maxbin2/), and checks the quality of the genome bins using [Busco](https://busco.ezlab.org/) -- optionally refines bins with [DAS_Tool](https://github.com/cmks/DAS_Tool) +- optionally refines bins with [DAS Tool](https://github.com/cmks/DAS_Tool) - assigns taxonomy to bins using [GTDB-Tk](https://github.com/Ecogenomics/GTDBTk) and/or [CAT](https://github.com/dutilh/CAT) Furthermore, the pipeline creates various reports in the results directory specified, including a [MultiQC](https://multiqc.info/) report summarizing some of the findings and software versions. diff --git a/docs/usage.md b/docs/usage.md index 0c6a4e72..7ee0a845 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -290,12 +290,12 @@ MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assem ## A note on bin refinement -DAS_Tool may not always be able to refine bins due to insufficient recovery of enough single-copy genes. In these cases you will get a NOTE such as +DAS Tool may not always be able to refine bins due to insufficient recovery of enough single-copy genes. In these cases you will get a NOTE such as ```console [16/d330a6] NOTE: Process `NFCORE_MAG:MAG:BINNING_REFINEMENT:DASTOOL_DASTOOL (test_minigut_sample2)` terminated with an error exit status (1) -- Error is ignored ``` -In this case, DAS_Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the BinRefinement results directory for that sample. +In this case, DAS Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the `GenomeBinningRefinement/` results directory for that sample. -If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS_Tool publication](https://www.nature.com/articles/s41564-018-0171-1). +If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS Tool publication](https://www.nature.com/articles/s41564-018-0171-1). diff --git a/modules/local/combine_tsv.nf b/modules/local/combine_tsv.nf index 789e4856..4564cd29 100644 --- a/modules/local/combine_tsv.nf +++ b/modules/local/combine_tsv.nf @@ -10,8 +10,8 @@ process COMBINE_TSV { path(bin_summaries) output: - path("*.tsv"), emit: combined - path "versions.yml" , emit: versions + path("*.tsv") , emit: combined + path "versions.yml", emit: versions script: def prefix = task.ext.prefix ?: "bin_depths_summary_combined" diff --git a/modules/local/mag_depths_summary.nf b/modules/local/mag_depths_summary.nf index a3c9e43f..75b7d963 100644 --- a/modules/local/mag_depths_summary.nf +++ b/modules/local/mag_depths_summary.nf @@ -10,7 +10,7 @@ process MAG_DEPTHS_SUMMARY { output: path("${prefix}.tsv"), emit: summary - path "versions.yml" , emit: versions + path "versions.yml" , emit: versions script: prefix = task.ext.prefix ?: "bin_depths_summary" diff --git a/modules/local/rename_postdastool.nf b/modules/local/rename_postdastool.nf index 8691b525..e5a69d65 100644 --- a/modules/local/rename_postdastool.nf +++ b/modules/local/rename_postdastool.nf @@ -12,7 +12,7 @@ process RENAME_POSTDASTOOL { tuple val(meta), path(bins) output: - tuple val(meta), path("${meta.assembler}-DASToolUnbinned-${meta.id}.fa") , optional:true, emit: refined_unbins + tuple val(meta), path("${meta.assembler}-DASToolUnbinned-${meta.id}.fa"), optional:true, emit: refined_unbins script: """ diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index d25cf902..919046b9 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -12,7 +12,7 @@ process RENAME_PREDASTOOL { tuple val(meta), path(bins) output: - tuple val(meta), path("${meta.assembler}-${meta.binner}Refined-${meta.id}*") , emit: renamed_bins + tuple val(meta), path("${meta.assembler}-${meta.binner}Refined-${meta.id}*"), emit: renamed_bins script: """ diff --git a/nextflow_schema.json b/nextflow_schema.json index ed5f9bff..9cdaf1ec 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -608,19 +608,19 @@ }, "refine_bins_dastool": { "type": "boolean", - "description": "Turn on bin refinement using DAS_Tool." + "description": "Turn on bin refinement using DAS Tool." }, "refine_bins_dastool_threshold": { "type": "number", "default": 0.5, "description": "Specify single-copy gene score threshold for bin refinement.", - "help_text": "Score threshold for single-copy gene selection algorithm to keep selecting bins, with a value ranging from 0-1.\n\nFor description of scoring algorithm, see: Sieber, Christian M. K., et al. 2018. Nature Microbiology 3 (7): 836\u201343. https://doi.org/10.1038/s41564-018-0171-1.\n\n> Modifies DAS_Tool parameter --score_threshold\n" + "help_text": "Score threshold for single-copy gene selection algorithm to keep selecting bins, with a value ranging from 0-1.\n\nFor description of scoring algorithm, see: Sieber, Christian M. K., et al. 2018. Nature Microbiology 3 (7): 836\u201343. https://doi.org/10.1038/s41564-018-0171-1.\n\n> Modifies DAS Tool parameter --score_threshold\n" }, "postbinning_input": { "type": "string", "default": "raw_bins_only", - "description": "Specify which binning output is sent for downstream annotation, taxonomic categorisation, assembly qualiy control etc.", - "help_text": "`raw_bins_only`: only bins (and unbinned contigs) from the binners.\n`refined_bins_only`: only bins (and unbinned contigs) from the bin refinement step .\n`both`: bins and and unbinned contigs from both the binning and bin refinement steps.", + "description": "Specify which binning output is sent for downstream annotation, taxonomic classification, bin quality control etc.", + "help_text": "`raw_bins_only`: only bins (and unbinned contigs) from the binners.\n`refined_bins_only`: only bins (and unbinned contigs) from the bin refinement step .\n`both`: bins and unbinned contigs from both the binning and bin refinement steps.", "enum": ["raw_bins_only", "refined_bins_only", "both"] } } diff --git a/workflows/mag.nf b/workflows/mag.nf index ee59e5d5..42d26dd3 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -551,7 +551,7 @@ workflow MAG { ch_versions = ch_versions.mix(BINNING.out.versions) /* - * DAS_Tool: binning refinement + * DAS Tool: binning refinement */ if ( params.refine_bins_dastool && !params.skip_metabat2 && !params.skip_maxbin2 ) { From 5e7556ce551f03cea5bebdf70f25db6ab8fa0385 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 13:45:40 +0200 Subject: [PATCH 188/246] Update output.md --- docs/output.md | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/docs/output.md b/docs/output.md index b8ac15fc..ac81980a 100644 --- a/docs/output.md +++ b/docs/output.md @@ -335,13 +335,13 @@ Files in these two folders contain all contigs of an assembly. Output files - `GenomeBinning/DASTool/` - - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets - - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets - - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners - - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS_Tool run describing the command executed and additional runtime information - - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig - - `bins/[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format - - `bins/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format + - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets. Quality and completeness are estimated by DAS TOOL using a scoring function based on the frequency of bacterial or archaeal reference single-copy genes (SCG). + - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets. + - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners. + - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS Tool run describing the command executed and additional runtime information. + - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig. + - `bins/[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format. + - `bins/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format.
@@ -354,7 +354,7 @@ For each genome bin the median sequencing depth is computed based on the corresp - `GenomeBinning/depths/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - - `bin_depths_summary_refined.tsv`: Summary of bin sequencing depths for all samples against refined bin contigs, if refinement performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. + - `bin_depths_summary_refined.tsv`: Summary of sequencing depths for refined bins for all samples, if refinement was performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. From 57fffae75728ee8b2be232782a7e31b3f8ac4168 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 14:18:58 +0200 Subject: [PATCH 189/246] Fix binning refinement output folder in usage.md --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index 7ee0a845..ce8064ad 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -296,6 +296,6 @@ DAS Tool may not always be able to refine bins due to insufficient recovery of e [16/d330a6] NOTE: Process `NFCORE_MAG:MAG:BINNING_REFINEMENT:DASTOOL_DASTOOL (test_minigut_sample2)` terminated with an error exit status (1) -- Error is ignored ``` -In this case, DAS Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the `GenomeBinningRefinement/` results directory for that sample. +In this case, DAS Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the `GenomeBinning/DASTool/` results directory for that sample. If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS Tool publication](https://www.nature.com/articles/s41564-018-0171-1). From 6a40c6013a3ecada026ae6c56ccc98dce45dced6 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 14:34:11 +0200 Subject: [PATCH 190/246] Fix link in output.md --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index ac81980a..5697f296 100644 --- a/docs/output.md +++ b/docs/output.md @@ -14,7 +14,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d - [Taxonomic classification of trimmed reads](#taxonomic-classification-of-trimmed-reads) - [Assembly](#assembly) of trimmed reads - [Protein-coding gene prediction](#gene-prediction) of assemblies -- [Binning](#binning) of assembled contigs +- [Binning and binning refinement](#binning-and-binning-refinement) of assembled contigs - [Taxonomic classification of binned genomes](#taxonomic-classification-of-binned-genomes) - [Genome annotation of binned genomes](#genome-annotation-of-binned-genomes) - [Additional summary for binned genomes](#additional-summary-for-binned-genomes) @@ -252,7 +252,7 @@ Protein-coding genes are predicted for each assembly. -## Binning and Binning Refinement +## Binning and binning refinement ### Contig sequencing depth From f011a444c847f2f436507a32cfd01e1dfdb71405 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 14:45:59 +0200 Subject: [PATCH 191/246] Extend documentation with respect to --postbinning_input --- docs/output.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 5697f296..fa905849 100644 --- a/docs/output.md +++ b/docs/output.md @@ -345,6 +345,8 @@ Files in these two folders contain all contigs of an assembly. +By default, only the raw bins (and unbinned contigs) from the actual binning methods, but not from the binning refinement with DAS Tool, will be used for downstream bin quality control, annotation and taxonomic classification. The parameter `--postbinning_input` can be used to change this behaviour. + ### Bin sequencing depth For each genome bin the median sequencing depth is computed based on the corresponding contig depths. @@ -386,7 +388,7 @@ For each genome bin the median sequencing depth is computed based on the corresp ### QC for metagenome assembled genomes with BUSCO -[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and high quality contigs obtained by the applied binning and binning refinement methods. By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated. +[BUSCO](https://busco.ezlab.org/) is a tool used to assess the completeness of a genome assembly. It is run on all the genome bins and high quality contigs obtained by the applied binning and/or binning refinement methods (depending on the `--postbinning_input` parameter). By default, BUSCO is run in automated lineage selection mode in which it first tries to select the domain and then a more specific lineage based on phylogenetic placement. If available, result files for both the selected domain lineage and the selected more specific lineage are placed in the output directory. If a lineage dataset is specified already with `--busco_reference`, only results for this specific lineage will be generated.
Output files From c7675ac49fb5ae38794a65ceb44c75004b9271e9 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 17 May 2022 21:28:10 +0200 Subject: [PATCH 192/246] Fix undefined channels and restructure code --- subworkflows/local/binning.nf | 35 +++++++++++++---------------------- 1 file changed, 13 insertions(+), 22 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 6dd5b117..7f4211c4 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -80,14 +80,22 @@ workflow BINNING { ch_versions = ch_versions.mix(CONVERT_DEPTHS.out.versions.first()) } + // main bins for decompressing for MAG_DEPTHS + ch_final_bins_for_gunzip = Channel.empty() + // final gzipped bins + ch_binning_results_gzipped_final = Channel.empty() // run binning if ( !params.skip_metabat2 ) { METABAT2_METABAT2 ( ch_metabat2_input ) + // before decompressing first have to separate and re-group due to limitation of GUNZIP module + ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( METABAT2_METABAT2.out.fasta.transpose() ) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( METABAT2_METABAT2.out.fasta ) ch_versions = ch_versions.mix(METABAT2_METABAT2.out.versions.first()) } - if ( !params.skip_maxbin2 ) { MAXBIN2 ( ch_maxbin2_input ) + ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( MAXBIN2.out.binned_fastas.transpose() ) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( MAXBIN2.out.binned_fastas ) ch_versions = ch_versions.mix(MAXBIN2.out.versions.first()) } @@ -101,25 +109,11 @@ workflow BINNING { } SPLIT_FASTA ( ch_input_splitfasta ) - - // decompress main bins (and large unbinned contigs from SPLIT_FASTA) for - // MAG_DEPTHS, first have to separate and re-group due to limitation of - // GUNZIP module - if ( !params.skip_metabat2 ) { - ch_metabat2_results_transposed = METABAT2_METABAT2.out.fasta.transpose() - } - - if ( !params.skip_maxbin2 ) { - ch_maxbin2_results_transposed = MAXBIN2.out.binned_fastas.transpose() - } - + // large unbinned contigs from SPLIT_FASTA for decompressing for MAG_DEPTHS, + // first have to separate and re-group due to limitation of GUNZIP module ch_split_fasta_results_transposed = SPLIT_FASTA.out.unbinned.transpose() ch_versions = ch_versions.mix(SPLIT_FASTA.out.versions) - // mix all bins together - ch_final_bins_for_gunzip = ch_metabat2_results_transposed - .mix( ch_maxbin2_results_transposed ) - GUNZIP_BINS ( ch_final_bins_for_gunzip ) ch_binning_results_gunzipped = GUNZIP_BINS.out.gunzip ch_versions = ch_versions.mix(GUNZIP_BINS.out.versions.first()) @@ -165,11 +159,8 @@ workflow BINNING { ch_versions = ch_versions.mix( MAG_DEPTHS_SUMMARY.out.versions ) // Group final binned contigs per sample for final output - ch_binning_results_gunzipped_final = ch_binning_results_gunzipped - .groupTuple(by: 0) - - ch_binning_results_gzipped_final = METABAT2_METABAT2.out.fasta.mix(MAXBIN2.out.binned_fastas) - .groupTuple(by: 0) + ch_binning_results_gunzipped_final = ch_binning_results_gunzipped.groupTuple(by: 0) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.groupTuple(by: 0) SPLIT_FASTA.out.unbinned From 3f25a8e94419e6c3d32b957a9f9db456353f4c33 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 19 May 2022 16:25:03 +0200 Subject: [PATCH 193/246] Fix wrong amount of left-padding spaces --- conf/modules.config | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index df449e2a..a389628b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -252,12 +252,12 @@ process { } withName: 'BUSCO_SUMMARY|QUAST_BINS|QUAST_BINS_SUMMARY' { - publishDir = [ - path: { "${params.outdir}/GenomeBinning/QC" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] - } + publishDir = [ + path: { "${params.outdir}/GenomeBinning/QC" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } withName: CAT_DB_GENERATE { publishDir = [ From ed20591abb0335bbce8f9367e29fabece611d5c7 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 20 May 2022 11:21:09 +0200 Subject: [PATCH 194/246] Fix published output for DAS TOOL --- conf/modules.config | 21 +++++++-------------- docs/output.md | 2 +- modules/local/rename_postdastool.nf | 3 ++- 3 files changed, 10 insertions(+), 16 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index a389628b..6f79ca0e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -471,29 +471,22 @@ process { ext.args = "--write_bins --write_unbinned --write_bin_evals --score_threshold ${params.refine_bins_dastool_threshold}" } - withName: RENAME_PREDASTOOL { - publishDir = [ - [ - path: { "${params.outdir}/GenomeBinning/DASTool/bins" }, - mode: params.publish_dir_mode, - pattern: '*.fa' - ], - ] - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } - } - withName: RENAME_POSTDASTOOL { publishDir = [ [ path: { "${params.outdir}/GenomeBinning/DASTool/unbinned" }, mode: params.publish_dir_mode, - pattern: '*.fa' + pattern: '*-DASToolUnbinned-*.fa' ], + [ + path: { "${params.outdir}/GenomeBinning/DASTool/bins" }, + mode: params.publish_dir_mode, + // pattern needs to be updated in case of new binning methods + pattern: '*-{MetaBAT2,MaxBin2}Refined-*.fa' + ] ] - ext.prefix = { "${meta.assembler}-${meta.binner}-${meta.id}" } } - withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/docs/output.md b/docs/output.md index fa905849..25586843 100644 --- a/docs/output.md +++ b/docs/output.md @@ -341,7 +341,7 @@ Files in these two folders contain all contigs of an assembly. - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS Tool run describing the command executed and additional runtime information. - `[assembler]-[sample/group].seqlength`: Tab-delimited file describing the length of each contig. - `bins/[assembler]-[binner]Refined-[sample/group].*.fa`: Refined bins in fasta format. - - `bins/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format. + - `unbinned/[assembler]-DASToolUnbinned-[sample/group].*.fa`: Unbinned contigs from bin refinement in fasta format.
diff --git a/modules/local/rename_postdastool.nf b/modules/local/rename_postdastool.nf index e5a69d65..7c746d01 100644 --- a/modules/local/rename_postdastool.nf +++ b/modules/local/rename_postdastool.nf @@ -12,7 +12,8 @@ process RENAME_POSTDASTOOL { tuple val(meta), path(bins) output: - tuple val(meta), path("${meta.assembler}-DASToolUnbinned-${meta.id}.fa"), optional:true, emit: refined_unbins + tuple val(meta), path("${meta.assembler}-*Refined-${meta.id}.*.fa", includeInputs: true), optional:true, emit: refined_bins + tuple val(meta), path("${meta.assembler}-DASToolUnbinned-${meta.id}.fa"), optional:true, emit: refined_unbins script: """ From 501c154969850924680c11abe1e61937c1ee8252 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 21 May 2022 11:06:47 +0200 Subject: [PATCH 195/246] Fix MaxBin2 extension expected for DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 --- subworkflows/local/binning_refinement.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index a0e666c3..62b1d504 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -45,7 +45,8 @@ workflow BINNING_REFINEMENT { // Generate DASTool auxilary files DASTOOL_FASTATOCONTIG2BIN_METABAT2 ( ch_bins_for_fastatocontig2bin.metabat2, "fa") - DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 ( ch_bins_for_fastatocontig2bin.maxbin2, "fasta") + // MaxBin2 bin extension was changed to 'fa' as well in RENAME_PREDASTOOL + DASTOOL_FASTATOCONTIG2BIN_MAXBIN2 ( ch_bins_for_fastatocontig2bin.maxbin2, "fa") // Run DASTOOL ch_fastatocontig2bin_for_dastool = Channel.empty() From e100186990805265d6b2ee6c3abdcb60c2008975 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 21 May 2022 11:46:39 +0200 Subject: [PATCH 196/246] Fix: separate BINNING_REFINEMENT refined_bins output from unbins --- subworkflows/local/binning_refinement.nf | 10 ++++++---- 1 file changed, 6 insertions(+), 4 deletions(-) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 62b1d504..744b6069 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -69,13 +69,15 @@ workflow BINNING_REFINEMENT { // Run DAStool DASTOOL_DASTOOL(ch_input_for_dastool, [], []) + // Prepare bins for downstream analysis (separate from unbins, add 'binner' info and group) ch_dastool_bins_newmeta = DASTOOL_DASTOOL.out.bins.transpose() .map { meta, bin -> - def meta_new = meta.clone() - - meta_new['binner'] = bin.name.split("-")[1] - [ meta_new, bin ] + if (bin.name != "unbinned.fa") { + def meta_new = meta.clone() + meta_new['binner'] = bin.name.split("-")[1] + [ meta_new, bin ] + } } .groupTuple() From d469e61f072f1617bbda77442739358da6ac9789 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 21 May 2022 12:19:38 +0200 Subject: [PATCH 197/246] Use DASTool as meta.binner info --- modules/local/busco_plot.nf | 6 +++--- subworkflows/local/binning_refinement.nf | 12 ++++++++---- 2 files changed, 11 insertions(+), 7 deletions(-) diff --git a/modules/local/busco_plot.nf b/modules/local/busco_plot.nf index c3ff6a8e..42392f27 100644 --- a/modules/local/busco_plot.nf +++ b/modules/local/busco_plot.nf @@ -21,14 +21,14 @@ process BUSCO_PLOT { # replace dots in bin names within summary file names by underscores # currently (BUSCO v5.1.0) generate_plot.py does not allow further dots for sum in ${summaries}; do - if [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt ]]; then + if [[ \${sum} =~ short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-([_[:alnum:]]+)-${meta.id}.(.+).txt ]]; then mode=\${BASH_REMATCH[1]} db_name=\${BASH_REMATCH[2]} - bin="${meta.assembler}-${meta.binner}-${meta.id}.\${BASH_REMATCH[3]}" + bin="${meta.assembler}-\${BASH_REMATCH[3]}-${meta.id}.\${BASH_REMATCH[4]}" bin_new="\${bin//./_}" mv \${sum} short_summary.\${mode}.\${db_name}.\${bin_new}.txt else - echo "ERROR: the summary filename \${sum} does not match the expected format 'short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-${meta.binner}-${meta.id}.(.+).txt'!" + echo "ERROR: the summary filename \${sum} does not match the expected format 'short_summary.([_[:alnum:]]+).([_[:alnum:]]+).${meta.assembler}-([_[:alnum:]]+)-${meta.id}.(.+).txt'!" exit 1 fi done diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 744b6069..85c5e517 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -68,21 +68,23 @@ workflow BINNING_REFINEMENT { // Run DAStool DASTOOL_DASTOOL(ch_input_for_dastool, [], []) + ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions.first()) // Prepare bins for downstream analysis (separate from unbins, add 'binner' info and group) + // use DASTool as 'binner' info allowing according grouping of refined bin sets, + // while keeping information about original binning method in filenames and used binnames, e.g. "*-MaxBin2Refined-*.fa" + // (alternatively one could think of adding, for example, meta.orig_binner, if this would simplify code) ch_dastool_bins_newmeta = DASTOOL_DASTOOL.out.bins.transpose() .map { meta, bin -> if (bin.name != "unbinned.fa") { def meta_new = meta.clone() - meta_new['binner'] = bin.name.split("-")[1] + meta_new['binner'] = 'DASTool' [ meta_new, bin ] } } .groupTuple() - ch_versions = ch_versions.mix(DASTOOL_DASTOOL.out.versions.first()) - ch_input_for_renamedastool = DASTOOL_DASTOOL.out.bins .map { meta, bins -> @@ -103,6 +105,8 @@ workflow BINNING_REFINEMENT { meta_new.remove('binner') [ meta_new, refinedbins ] } + .transpose() + .groupTuple (by: 0 ) .join( depths, by: 0 ) MAG_DEPTHS_REFINED ( ch_input_for_magdepth ) @@ -118,7 +122,7 @@ workflow BINNING_REFINEMENT { .transpose() .map { meta, depth_file -> def meta_new = meta.clone() - meta_new['binner'] = depth_file.name.split("-")[1] + meta_new['binner'] = 'DASTool' if (getColNo(depth_file) > 2) [meta_new, depth_file] } From f48b83a003ee998b53c891e0200905156bb662a7 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 23 May 2022 13:54:05 +0200 Subject: [PATCH 198/246] Run MAG_DEPTHS on each meta.binner, adjust output filenames and group input channels accordingly --- bin/get_mag_depths.py | 10 ++++----- modules/local/mag_depths.nf | 9 ++++---- subworkflows/local/binning.nf | 26 ++++++++++++++---------- subworkflows/local/binning_refinement.nf | 16 ++++++++------- 4 files changed, 34 insertions(+), 27 deletions(-) diff --git a/bin/get_mag_depths.py b/bin/get_mag_depths.py index 9a13f591..64418068 100755 --- a/bin/get_mag_depths.py +++ b/bin/get_mag_depths.py @@ -17,7 +17,9 @@ def parse_args(args=None): parser.add_argument('-d', '--depths' , required=True , metavar='FILE' , help="(Compressed) TSV file containing contig depths for each sample: contigName, contigLen, totalAvgDepth, sample1_avgDepth, sample1_var [, sample2_avgDepth, sample2_var, ...].") parser.add_argument('-a', '--assembler' , required=True , type=str , help="Assembler name.") parser.add_argument('-i', '--id' , required=True , type=str , help="Sample or group id.") + parser.add_argument('-m', '--binner' , required=True , type=str , help="Binning method.") return parser.parse_args(args) +# Processing contig depths for each binner again, i.e. not the most efficient way, but ok def main(args=None): args = parse_args(args) @@ -43,10 +45,8 @@ def main(args=None): # Initialize output files n_samples = len(sample_names) - binners = set([ os.path.basename(file).split("-")[1] for file in args.bins ]) - for binner in binners: - with open(args.assembler + "-" + binner + "-" + args.id + "-binDepths.tsv", 'w') as outfile: - print("bin", '\t'.join(sample_names), sep='\t', file=outfile) + with open(args.assembler + "-" + args.binner + "-" + args.id + "-binDepths.tsv", 'w') as outfile: + print("bin", '\t'.join(sample_names), sep='\t', file=outfile) # for each bin, access contig depths and compute mean bin depth (for all samples) for file in args.bins: @@ -66,7 +66,7 @@ def main(args=None): all_depths[sample].append(contig_depths[sample]) binname = os.path.basename(file) - with open(args.assembler + "-" + binname.split("-")[1] + "-" + args.id + "-binDepths.tsv", 'a') as outfile: + with open(args.assembler + "-" + args.binner + "-" + args.id + "-binDepths.tsv", 'a') as outfile: print(binname, '\t'.join(str(statistics.median(sample_depths)) for sample_depths in all_depths), sep='\t', file=outfile) diff --git a/modules/local/mag_depths.nf b/modules/local/mag_depths.nf index b7e09f23..5a54385a 100644 --- a/modules/local/mag_depths.nf +++ b/modules/local/mag_depths.nf @@ -1,5 +1,5 @@ process MAG_DEPTHS { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" // Using container from metabat2 process, since this will be anyway already downloaded and contains biopython and pandas conda (params.enable_conda ? "bioconda::metabat2=2.15 conda-forge::python=3.6.7 conda-forge::biopython=1.74 conda-forge::pandas=1.1.5" : null) @@ -11,15 +11,16 @@ process MAG_DEPTHS { tuple val(meta), path(bins), path(contig_depths) output: - tuple val(meta), path("${meta.assembler}-*-${meta.id}-binDepths.tsv"), emit: depths - path "versions.yml" , emit: versions + tuple val(meta), path("${meta.assembler}-${meta.binner}-${meta.id}-binDepths.tsv"), emit: depths + path "versions.yml" , emit: versions script: """ get_mag_depths.py --bins ${bins} \\ --depths ${contig_depths} \\ --assembler ${meta.assembler} \\ - --id ${meta.id} + --id ${meta.id} \\ + --binner ${meta.binner} cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 7f4211c4..7f857cd2 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -123,17 +123,25 @@ workflow BINNING { ch_versions = ch_versions.mix(GUNZIP_UNBINS.out.versions.first()) // Compute bin depths for different samples (according to `binning_map_mode`) - // Have to remove binner meta for grouping to mix back with original depth - // files, as required for MAG_DEPTHS + // Have to remove binner meta before joining with according depths files, + // as required for MAG_DEPTHS + // add 'binner' info again and finally group by 'assembler', 'id', 'binner' ch_depth_input = ch_binning_results_gunzipped .mix(ch_splitfasta_results_gunzipped ) - .map { meta, results -> + .map { meta, bin -> def meta_new = meta.clone() meta_new.remove('binner') - [ meta_new, results ] + [ meta_new, bin ] } .groupTuple (by: 0 ) .join( METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS.out.depth, by: 0 ) + .transpose() + .map { meta, bin, contig_depths_file -> + def meta_new = meta.clone() + meta_new['binner'] = bin.name.split("-")[1] + [ meta_new, bin, contig_depths_file ] + } + .groupTuple (by: [0,2] ) MAG_DEPTHS ( ch_depth_input ) ch_versions = ch_versions.mix(MAG_DEPTHS.out.versions) @@ -143,14 +151,10 @@ workflow BINNING { ch_sample_groups = reads .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } - // Transpose and add 'binner' meta information again for plotting - // filter MAG depth files: use only those for plotting that contain depths for > 2 samples + // Filter MAG depth files: use only those for plotting that contain depths for > 2 samples ch_mag_depths_plot = MAG_DEPTHS.out.depths - .transpose() - .map { meta, depth_file -> - def meta_new = meta.clone() - meta_new['binner'] = depth_file.name.split("-")[1] - if (getColNo(depth_file) > 2) [ meta_new, depth_file ] + .map { meta, bin_depths_file -> + if (getColNo(bin_depths_file) > 2) [ meta, bin_depths_file ] } MAG_DEPTHS_PLOT ( ch_mag_depths_plot, ch_sample_groups.collect() ) diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 85c5e517..17276583 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -108,6 +108,12 @@ workflow BINNING_REFINEMENT { .transpose() .groupTuple (by: 0 ) .join( depths, by: 0 ) + .map { + meta, bins, contig_depths_file -> + def meta_new = meta.clone() + meta_new['binner'] = 'DASTool' + [ meta_new, bins, contig_depths_file ] + } MAG_DEPTHS_REFINED ( ch_input_for_magdepth ) @@ -116,14 +122,10 @@ workflow BINNING_REFINEMENT { ch_sample_groups = reads .collectFile(name:'sample_groups.tsv'){ meta, reads -> meta.id + '\t' + meta.group + '\n' } - // Transpose and add 'binner' meta information again for plotting - // filter MAG depth files: use only those for plotting that contain depths for > 2 samples + // Filter MAG depth files: use only those for plotting that contain depths for > 2 samples ch_mag_depths_plot_refined = MAG_DEPTHS_REFINED.out.depths - .transpose() - .map { meta, depth_file -> - def meta_new = meta.clone() - meta_new['binner'] = 'DASTool' - if (getColNo(depth_file) > 2) [meta_new, depth_file] + .map { meta, bin_depths_file -> + if (getColNo(bin_depths_file) > 2) [ meta, bin_depths_file ] } MAG_DEPTHS_PLOT_REFINED ( ch_mag_depths_plot_refined, ch_sample_groups.collect() ) From 0415bce7673fa3c28b9ce6e5074dd8fcde5173fc Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 23 May 2022 14:39:57 +0200 Subject: [PATCH 199/246] Change output filenames bin_depths_summary_refined.tsv -> bin_refined_depths_summary.tsv --- conf/modules.config | 2 +- docs/output.md | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 6f79ca0e..3c856808 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -210,7 +210,7 @@ process { } withName: 'MAG_DEPTHS_SUMMARY_REFINED' { - ext.prefix = "bin_depths_summary_refined" + ext.prefix = "bin_refined_depths_summary" publishDir = [ path: { "${params.outdir}/GenomeBinning/depths" }, mode: params.publish_dir_mode, diff --git a/docs/output.md b/docs/output.md index 25586843..b4d735d6 100644 --- a/docs/output.md +++ b/docs/output.md @@ -356,7 +356,7 @@ For each genome bin the median sequencing depth is computed based on the corresp - `GenomeBinning/depths/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - - `bin_depths_summary_refined.tsv`: Summary of sequencing depths for refined bins for all samples, if refinement was performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. + - `bin_refined_depths_summary.tsv`: Summary of sequencing depths for refined bins for all samples, if refinement was performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. From baa1aa64e259313444c46709f3f4e58ae5bc1562 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 25 May 2022 15:02:24 +0200 Subject: [PATCH 200/246] Update CHANGELOG.md Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- CHANGELOG.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index a3eb1a8d..9e33c3c6 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,8 +10,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Restructure binning subworkflow in preparation for aDNA workflow and extended binning - [#247](https://github.com/nf-core/mag/pull/247) - Add ancient DNA subworkflow - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool -- [#284](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer -- [#284](https://github.com/nf-core/mag/pull/291) - Add DAS Tool for bin refinement +- [#285](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer +- [#291](https://github.com/nf-core/mag/pull/291) - Add DAS Tool for bin refinement ### `Changed` From dd4d07ac093f3061caafbc9c432ede5f55ce69f1 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 27 May 2022 15:16:44 +0200 Subject: [PATCH 201/246] Add Bowtie2 version of BOWTIE2_PHIX_REMOVAL_ALIGN --- workflows/mag.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/mag.nf b/workflows/mag.nf index 616ca573..bec9ed5d 100644 --- a/workflows/mag.nf +++ b/workflows/mag.nf @@ -277,6 +277,7 @@ workflow MAG { BOWTIE2_PHIX_REMOVAL_BUILD.out.index ) ch_short_reads = BOWTIE2_PHIX_REMOVAL_ALIGN.out.reads + ch_versions = ch_versions.mix(BOWTIE2_PHIX_REMOVAL_ALIGN.out.versions.first()) } FASTQC_TRIMMED ( From f0c3cabf5a80ae096455f4ab96d7a04427d08bbb Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 27 May 2022 22:29:42 +0200 Subject: [PATCH 202/246] Fix description of MaxBin2 output filenames --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index 48b2abd5..664182e8 100644 --- a/docs/output.md +++ b/docs/output.md @@ -305,7 +305,7 @@ Files in these two folders contain all contigs of an assembly. Output files - `GenomeBinning/MaxBin2/` - - `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly + - `bins/[assembler]-[binner]-[sample/group].*.fasta.gz`: Genome bins retrieved from input assembly - `unbinned/[assembler]-[binner]-[sample/group].noclass.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. @@ -316,7 +316,7 @@ All the files and contigs in these folders will be assessed by QUAST and BUSCO. Output files - `GenomeBinning/MaxBin2/discarded/` - - `*.tooshort.fa.gz`: Too short contigs that are filtered by MaxBin2 + - `*.tooshort.gz`: Too short contigs that are filtered by MaxBin2 - `GenomeBinning/MaxBin2/unbinned/discarded/` - `*.noclass.pooled.fa.gz`: Pooled unbinned contigs equal or above `--min_contig_size`, by default 1500 bp. - `*.noclass.remaining.fa.gz`: Remaining unbinned contigs below `--min_contig_size`, by default 1500 bp, but not in any other file. From 20c27baad91caae7e642fac9e33cacf44e08de48 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 27 May 2022 22:13:11 +0200 Subject: [PATCH 203/246] Adjust MaxBin2 bin output file extension to fa --- conf/modules.config | 15 ++++++++++----- docs/output.md | 2 +- modules/local/adjust_maxbin2_ext.nf | 28 ++++++++++++++++++++++++++++ subworkflows/local/binning.nf | 6 ++++-- 4 files changed, 43 insertions(+), 8 deletions(-) create mode 100644 modules/local/adjust_maxbin2_ext.nf diff --git a/conf/modules.config b/conf/modules.config index 650e95d7..ad530de6 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -406,11 +406,6 @@ process { withName: MAXBIN2 { publishDir = [ - [ - path: { "${params.outdir}/GenomeBinning/MaxBin2/bins/" }, - mode: params.publish_dir_mode, - pattern: '*.*[0-9].fasta.gz' - ], [ path: { "${params.outdir}/GenomeBinning/MaxBin2/discarded" }, mode: params.publish_dir_mode, @@ -423,6 +418,16 @@ process { errorStrategy = { task.exitStatus in [ 1, 255 ] ? 'ignore' : 'retry' } } + withName: ADJUST_MAXBIN2_EXT { + publishDir = [ + [ + path: { "${params.outdir}/GenomeBinning/MaxBin2/bins/" }, + mode: params.publish_dir_mode, + pattern: '*.fa.gz' + ], + ] + } + withName: SPLIT_FASTA { publishDir = [ [ diff --git a/docs/output.md b/docs/output.md index 664182e8..ddd2aa9e 100644 --- a/docs/output.md +++ b/docs/output.md @@ -305,7 +305,7 @@ Files in these two folders contain all contigs of an assembly. Output files - `GenomeBinning/MaxBin2/` - - `bins/[assembler]-[binner]-[sample/group].*.fasta.gz`: Genome bins retrieved from input assembly + - `bins/[assembler]-[binner]-[sample/group].*.fa.gz`: Genome bins retrieved from input assembly - `unbinned/[assembler]-[binner]-[sample/group].noclass.[1-9]*.fa.gz`: Contigs that were not binned with other contigs but considered interesting. By default, these are at least 1 Mbp (`--min_length_unbinned_contigs`) in length and at most the 100 longest contigs (`--max_unbinned_contigs`) are reported. diff --git a/modules/local/adjust_maxbin2_ext.nf b/modules/local/adjust_maxbin2_ext.nf new file mode 100644 index 00000000..63f9a211 --- /dev/null +++ b/modules/local/adjust_maxbin2_ext.nf @@ -0,0 +1,28 @@ +process ADJUST_MAXBIN2_EXT { + tag "${meta.assembler}-${meta.id}" + label 'process_low' + + // Using container from multiqc since it'll be included anyway + conda (params.enable_conda ? "bioconda::multiqc=1.12" : null) + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.12--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.12--pyhdfd78af_0' }" + + input: + tuple val(meta), path(bins) + + output: + tuple val(meta), path("*.fa.gz"), emit: renamed_bins + + script: + """ + if [ -n "${bins}" ] + then + for file in ${bins}; do + [[ \${file} =~ (.*).fasta.gz ]]; + bin="\${BASH_REMATCH[1]}" + mv \${file} \${bin}.fa.gz + done + fi + """ +} diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 06f498b9..2a8042d7 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -13,6 +13,7 @@ include { GUNZIP as GUNZIP_BINS } from '../../modules/nf-core/mo include { GUNZIP as GUNZIP_UNBINS } from '../../modules/nf-core/modules/gunzip/main' include { CONVERT_DEPTHS } from '../../modules/local/convert_depths' +include { ADJUST_MAXBIN2_EXT } from '../../modules/local/adjust_maxbin2_ext' include { SPLIT_FASTA } from '../../modules/local/split_fasta' include { MAG_DEPTHS } from '../../modules/local/mag_depths' addParams( options: params.mag_depths_options ) include { MAG_DEPTHS_PLOT } from '../../modules/local/mag_depths_plot' addParams( options: params.mag_depths_plot_options ) @@ -93,8 +94,9 @@ workflow BINNING { } if ( !params.skip_maxbin2 ) { MAXBIN2 ( ch_maxbin2_input ) - ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( MAXBIN2.out.binned_fastas.transpose() ) - ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( MAXBIN2.out.binned_fastas ) + ADJUST_MAXBIN2_EXT ( MAXBIN2.out.binned_fastas ) + ch_final_bins_for_gunzip = ch_final_bins_for_gunzip.mix( ADJUST_MAXBIN2_EXT.out.renamed_bins.transpose() ) + ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.mix( ADJUST_MAXBIN2_EXT.out.renamed_bins ) ch_versions = ch_versions.mix(MAXBIN2.out.versions) } From 68a0ea4d05058ae9acf1be70d6094d40135a424b Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 28 May 2022 17:07:36 +0200 Subject: [PATCH 204/246] Fix parsing of GTDB-Tk version --- modules/local/gtdbtk_classify.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gtdbtk_classify.nf b/modules/local/gtdbtk_classify.nf index 0724d472..4ad1e72c 100644 --- a/modules/local/gtdbtk_classify.nf +++ b/modules/local/gtdbtk_classify.nf @@ -47,7 +47,7 @@ process GTDBTK_CLASSIFY { cat <<-END_VERSIONS > versions.yml "${task.process}": - gtdbtk: \$(gtdbtk --version | sed "s/gtdbtk: version //; s/ Copyright.*//") + gtdbtk: \$(gtdbtk --version | head -n 1 | sed "s/gtdbtk: version //; s/ Copyright.*//") END_VERSIONS """ } From 5da7d08dbfe0c2f12507eecb718a61a6d4024660 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 28 May 2022 21:47:35 +0200 Subject: [PATCH 205/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 0580ab0d..a832611a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -22,6 +22,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Fixed` - [#290](https://github.com/nf-core/mag/pull/290) - Fix caching of binning input +- [#307](https://github.com/nf-core/mag/pull/307) - Fix retrieval of GTDB-Tk version (note about newer version caused error in `CUSTOM_DUMPSOFTWAREVERSIONS`) ### `Dependencies` From 40fa130efe520c7f2155f70f15a165dc0184b6e8 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Sat, 28 May 2022 21:52:15 +0200 Subject: [PATCH 206/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 0580ab0d..011790ab 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -22,6 +22,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Fixed` - [#290](https://github.com/nf-core/mag/pull/290) - Fix caching of binning input +- 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s/ Copyright.*//") + gtdbtk: \$(gtdbtk --version | sed -n 1p | sed "s/gtdbtk: version //; s/ Copyright.*//") END_VERSIONS """ } From 75f5b0b210ef315c99806597b2e1928b645ec95a Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 30 May 2022 21:32:42 +0200 Subject: [PATCH 210/246] Fix publishing of BUSCO downloads --- conf/modules.config | 12 ++++++++++-- 1 file changed, 10 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index ad530de6..5b300e4b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -225,11 +225,11 @@ process { ] } - withName: 'BUSCO|BUSCO_PLOT' { + withName: 'BUSCO' { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + pattern: "*.{log,err,faa.gz,fna.gz,gff,txt}" ] } @@ -242,6 +242,14 @@ process { ] } + withName: 'BUSCO_PLOT' { + publishDir = [ + path: { "${params.outdir}/GenomeBinning/QC/BUSCO" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + withName: 'BUSCO_SUMMARY|QUAST_BINS|QUAST_BINS_SUMMARY' { publishDir = [ path: { "${params.outdir}/GenomeBinning/QC" }, From f4d30e20b7ad7ec545d559fbc38c62ee10623a7c Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 30 May 2022 21:48:06 +0200 Subject: [PATCH 211/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7ac53d73..ca8eb5f1 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,6 +24,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#290](https://github.com/nf-core/mag/pull/290) - Fix caching of binning input - [#305](https://github.com/nf-core/mag/pull/305) - Add missing Bowtie2 version for process `BOWTIE2_PHIX_REMOVAL_ALIGN` to `software_versions.yml` - [#307](https://github.com/nf-core/mag/pull/307) - Fix retrieval of GTDB-Tk version (note about newer version caused error in `CUSTOM_DUMPSOFTWAREVERSIONS`) +- [#309](https://github.com/nf-core/mag/pull/309) - Fix publishing of BUSCO `busco_downloads/` folder, i.e. publish only when `--save_busco_reference` is specified ### `Dependencies` From b5097d1708a79b4b0721d585fe47afda3373b0ec Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Tue, 31 May 2022 18:00:33 +0200 Subject: [PATCH 212/246] Update docs/usage.md Co-authored-by: James A. Fellows Yates --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index ce8064ad..c66a26fa 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -296,6 +296,6 @@ DAS Tool may not always be able to refine bins due to insufficient recovery of e [16/d330a6] NOTE: Process `NFCORE_MAG:MAG:BINNING_REFINEMENT:DASTOOL_DASTOOL (test_minigut_sample2)` terminated with an error exit status (1) -- Error is ignored ``` -In this case, DAS Tool pipeline has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the `GenomeBinning/DASTool/` results directory for that sample. +In this case, DAS Tool has not necessarily failed but was unable to complete the refinement. You will therefore not expect to find any output files in the `GenomeBinning/DASTool/` results directory for that particular sample. If you are regularly getting such errors, you can try reducing the `--refine_bins_dastool_threshold` value, which will modify the scoring threshold defined in the [DAS Tool publication](https://www.nature.com/articles/s41564-018-0171-1). From 2a98e605967fb0f5d184f860caadb8f507cd9de3 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 1 Jun 2022 15:11:30 +0200 Subject: [PATCH 213/246] Update docs/usage.md Co-authored-by: James A. Fellows Yates --- docs/usage.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index c66a26fa..b24e000f 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -290,6 +290,8 @@ MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assem ## A note on bin refinement +### Error Reporting + DAS Tool may not always be able to refine bins due to insufficient recovery of enough single-copy genes. In these cases you will get a NOTE such as ```console From b515528142745028fe024503c26e79ad0028ed97 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 1 Jun 2022 15:25:41 +0200 Subject: [PATCH 214/246] Tweak docs and output structure for depths files for clarity --- conf/modules.config | 6 +++--- docs/output.md | 12 ++++++++---- 2 files changed, 11 insertions(+), 7 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index da248a50..1eafc2fe 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -203,7 +203,7 @@ process { withName: 'MAG_DEPTHS_PLOT|MAG_DEPTHS_SUMMARY|MAG_DEPTHS_PLOT_REFINED' { publishDir = [ - path: { "${params.outdir}/GenomeBinning/depths" }, + path: { "${params.outdir}/GenomeBinning/depths/bins" }, mode: params.publish_dir_mode, pattern: "*.{png,tsv}" ] @@ -212,7 +212,7 @@ process { withName: 'MAG_DEPTHS_SUMMARY_REFINED' { ext.prefix = "bin_refined_depths_summary" publishDir = [ - path: { "${params.outdir}/GenomeBinning/depths" }, + path: { "${params.outdir}/GenomeBinning/depths/bins" }, mode: params.publish_dir_mode, pattern: "*.{tsv}" ] @@ -388,7 +388,7 @@ process { } withName: METABAT2_JGISUMMARIZEBAMCONTIGDEPTHS { publishDir = [ - path: { "${params.outdir}/GenomeBinning/depths" }, + path: { "${params.outdir}/GenomeBinning/depths/contigs" }, mode: params.publish_dir_mode, pattern: '*-depth.txt.gz' ] diff --git a/docs/output.md b/docs/output.md index b57511b4..fb5d2032 100644 --- a/docs/output.md +++ b/docs/output.md @@ -258,10 +258,12 @@ Protein-coding genes are predicted for each assembly. Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig_depths --outputDepth`. The values correspond to `(sum of exactly aligned bases) / ((contig length)-2*75)`. For example, for two reads aligned exactly with `10` and `9` bases on a 1000 bp long contig the depth is calculated by `(10+9)/(1000-2*75)` (1000bp length of contig minus 75bp from each end, which is excluded). +These depth files are used for downstream binning steps. +
Output files -- `GenomeBinning/depths/` +- `GenomeBinning/depths/contigs/` - `[assembler]-[sample/group]-depth.txt.gz`: Sequencing depth for each contig and sample or group, only for short reads.
@@ -335,7 +337,7 @@ Files in these two folders contain all contigs of an assembly. Output files - `GenomeBinning/DASTool/` - - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets. Quality and completeness are estimated by DAS TOOL using a scoring function based on the frequency of bacterial or archaeal reference single-copy genes (SCG). + - `[assembler]-[sample/group]_allBins.eval`: Tab-delimited description with quality and completeness metrics for the input bin sets. Quality and completeness are estimated by DAS TOOL using a scoring function based on the frequency of bacterial or archaeal reference single-copy genes (SCG). Please see note at the bottom of this section on file names. - `[assembler]-[sample/group]_DASTool_summary.tsv`: Tab-delimited description with quality and completeness metrics for the refined output bin sets. - `[assembler]-[sample/group]_DASTool_contig2bin.tsv`: File describing which contig is associated to which bin from the input binners. - `[assembler]-[sample/group]_DASTool.log`: Log file from the DAS Tool run describing the command executed and additional runtime information. @@ -347,14 +349,16 @@ Files in these two folders contain all contigs of an assembly. By default, only the raw bins (and unbinned contigs) from the actual binning methods, but not from the binning refinement with DAS Tool, will be used for downstream bin quality control, annotation and taxonomic classification. The parameter `--postbinning_input` can be used to change this behaviour. +⚠️ Due to ability to perform downstream QC of both raw and refined bins in parallel (via `--postbinning_input)`, bin names in DAS Tools's `*_allBins.eval` file will include `Refined`, however for this particular file, they _actually_ refer to the 'raw' input bins. The pipeline renames the input files prior to running DASTool to ensure they can be disambuguated from the original bin files in the downstream QC steps. + ### Bin sequencing depth -For each genome bin the median sequencing depth is computed based on the corresponding contig depths. +For each genome bin the median sequencing depth is computed based on the corresponding contig depths of each bin or refined bin.
Output files -- `GenomeBinning/depths/` +- `GenomeBinning/depths/bins/` - `bin_depths_summary.tsv`: Summary of bin sequencing depths for all samples. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `bin_refined_depths_summary.tsv`: Summary of sequencing depths for refined bins for all samples, if refinement was performed. Depths are available for samples mapped against the corresponding assembly, i.e. according to the mapping strategy specified with `--binning_map_mode`. Only for short reads. - `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png`: Clustered heatmap showing bin abundances of the assembly across samples. Bin depths are transformed to centered log-ratios and bins as well as samples are clustered by Euclidean distance. Again, sample depths are available according to the mapping strategy specified with `--binning_map_mode`. From a433d97a4c1e9088cfba0f790fd985e6004f1380 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 1 Jun 2022 15:54:38 +0200 Subject: [PATCH 215/246] Remove re-enumeration of bin files for DAS Tool --- modules/local/rename_predastool.nf | 17 ++++++++++++----- 1 file changed, 12 insertions(+), 5 deletions(-) diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index 919046b9..af670f4e 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -16,10 +16,17 @@ process RENAME_PREDASTOOL { script: """ - SAMPLES=(*) - - for i in \$(seq 0 \$((\${#SAMPLES[@]}-1))); do - mv \${SAMPLES[\$i]} ${meta.assembler}-${meta.binner}Refined-${meta.id}.\$(( \$i + 1)).fa - done + if [ -n "${bins}" ] + then + for bin in ${bins}; do + if [[ \${bin} =~ ${meta.assembler}-${meta.binner}-${meta.id}.([_[:alnum:]]+).fa ]]; then + num=\${BASH_REMATCH[1]} + mv \${bin} ${meta.assembler}-${meta.binner}Refined-${meta.id}.\${num}.fa + else + echo "ERROR: the bin filename \${bin} does not match the expected format '${meta.assembler}-${meta.binner}-${meta.id}.([_[:alnum:]]+).fa'!" + exit 1 + fi + done + fi """ } From 0b758aee09677f5ae800595a61fd2dc5d82e1415 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Wed, 1 Jun 2022 20:30:15 +0200 Subject: [PATCH 216/246] Add missing components to workflow diagram and polishing everywhere --- docs/images/mag_workflow.svg | 712 ++++++++++++++++--------------- docs/images/mag_workflow_new.png | Bin 0 -> 588955 bytes 2 files changed, 378 insertions(+), 334 deletions(-) create mode 100644 docs/images/mag_workflow_new.png diff --git a/docs/images/mag_workflow.svg b/docs/images/mag_workflow.svg index 8c553fdc..22cd66cc 100644 --- a/docs/images/mag_workflow.svg +++ b/docs/images/mag_workflow.svg @@ -1,23 +1,23 @@ + inkscape:export-ydpi="289.40701" + xmlns:inkscape="http://www.inkscape.org/namespaces/inkscape" + xmlns:sodipodi="http://sodipodi.sourceforge.net/DTD/sodipodi-0.dtd" + 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z-B==RumdQ@ntczdSLalbR(j1CA5R0g@)7gFmzxJbj6fhv=g{<)du}I3_@)Ex&%6M;{)Mv|9TaMtJmuoYPaIO6BmF6 zMr+T&&CGUU`^hwoVDt`{qo5KSkTgOfS4RE0|DYIUqn>W|> zdskqN;bvN3A-4|7Mgyu?u&*Q_sS4}Q2pEf~l;funNJrr}!nWp1ZTvb1{dpw|fh{5THk-s=-mr0_+PgwI;!UkS!dfm?v$mCXqejN>Dqf^iP7E;JQ$hY|CuY%Q z8njT|#q#HM75RIl3W!iq4{|y{Jo?^6V5RCLj8rO-NLV{;Dz!hWp+j?DwtXvy=XFGu z6_Biw@>wz=Rn3kVt?YAet}FjhJ;XVpG`bt8Mhj=B4XY&yH~n@(pBe%r5ejN{QyHIyG_ z&ew4L_Zh9&2bDt(oV$0=ZTqZ!+k0-iFzCdQjgMA-a z+mMlw5oP^JRvp*4Fr3{ctE*i(*Z5ppTS$}Ho55K**qzV`IgfAaLHTWILGw)VA6R&U<;hk`e%cFg+PXuvP6t%1%iE>mtEd-U?9 zYiJ*Z;y0U+Q?7@O969fAy6KL8zki+4^YHZaz0v`*xJ1qRu6muG)nPq9=VkQ$u_bQf zb*5GXUJG4_3`~7?6<>ghWzxiy3Pr}}*$y>H=oi3u& z%;_8I>gra8gnU2iVgVx&Xk^o}tfi%8l!-}S8|66;(;aSLP!=aEOZ~E#^;QRQ^yd%s z_P92^8NrSp=JjH!{{5w5T>t+6VF5gpzrQTT{_m@5Gd_QRMQq{3x4*xpO;xi!fBh(7 zVAy{zjglV!|26&^p8p?>zs7>5u>aF@qovzBSFT6-f1 Date: Thu, 2 Jun 2022 10:18:53 +0200 Subject: [PATCH 218/246] Update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c0c1b275..cdeecb21 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,7 +17,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289) - Update to nf-core 2.3.2 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint -- [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths` for `[assembler]-[sample/group]-depth.txt.gz`, `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` +- [#291](https://github.com/nf-core/mag/pull/291), [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths/contigs` for `[assembler]-[sample/group]-depth.txt.gz`, and `GenomeBinning/depths/bins` for `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` - [#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` ### `Fixed` From b5d5da980cea1ea8552a6eaad3084e58f9979a4a Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Thu, 2 Jun 2022 10:19:18 +0200 Subject: [PATCH 219/246] Bump version 2.2.0 --- CHANGELOG.md | 2 +- nextflow.config | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index cdeecb21..5f843a84 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v2.2.0dev +## v2.2.0 - 2022/06/02 ### `Added` diff --git a/nextflow.config b/nextflow.config index 867d228e..789673e2 100644 --- a/nextflow.config +++ b/nextflow.config @@ -267,7 +267,7 @@ manifest { description = 'Assembly, binning and annotation of metagenomes' mainScript = 'main.nf' nextflowVersion = '!>=21.10.3' - version = '2.2.0dev' + version = '2.2.0' } // Load modules.config for DSL2 module specific options From a5dc5cc49c814b090375ebede7b62188d728389b Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Jun 2022 08:44:50 +0200 Subject: [PATCH 220/246] Replace busybox with Ubuntu base image for GCP support --- modules.json | 2 +- modules/local/busco_db_preparation.nf | 4 ++-- modules/local/busco_save_download.nf | 4 ++-- modules/local/centrifuge_db_preparation.nf | 4 ++-- modules/local/gtdbtk_db_preparation.nf | 4 ++-- modules/local/kraken2_db_preparation.nf | 4 ++-- modules/local/pool_paired_reads.nf | 4 ++-- modules/local/pool_single_reads.nf | 4 ++-- modules/local/quast_bins_summary.nf | 4 ++-- modules/nf-core/modules/gunzip/main.nf | 4 ++-- 10 files changed, 19 insertions(+), 19 deletions(-) diff --git a/modules.json b/modules.json index f09b78c5..c1f58a4d 100644 --- a/modules.json +++ b/modules.json @@ -34,7 +34,7 @@ "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" }, "gunzip": { - "git_sha": "e745e167c1020928ef20ea1397b6b4d230681b4d" + "git_sha": "9aadd9a6d3f5964476582319b3a1c54a3e3fe7c9" }, "maxbin2": { "git_sha": "b78a4a456762a4c59fd5023e70f36a27f76d4a97" diff --git a/modules/local/busco_db_preparation.nf b/modules/local/busco_db_preparation.nf index 9b8de4e3..9d655a02 100644 --- a/modules/local/busco_db_preparation.nf +++ b/modules/local/busco_db_preparation.nf @@ -3,8 +3,8 @@ process BUSCO_DB_PREPARATION { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path database diff --git a/modules/local/busco_save_download.nf b/modules/local/busco_save_download.nf index 839cc7da..63928f85 100644 --- a/modules/local/busco_save_download.nf +++ b/modules/local/busco_save_download.nf @@ -4,8 +4,8 @@ process BUSCO_SAVE_DOWNLOAD { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path(busco_downloads) diff --git a/modules/local/centrifuge_db_preparation.nf b/modules/local/centrifuge_db_preparation.nf index 2f1f7c04..f1a99afe 100644 --- a/modules/local/centrifuge_db_preparation.nf +++ b/modules/local/centrifuge_db_preparation.nf @@ -2,8 +2,8 @@ process CENTRIFUGE_DB_PREPARATION { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path db diff --git a/modules/local/gtdbtk_db_preparation.nf b/modules/local/gtdbtk_db_preparation.nf index f2d79053..aabd1e0a 100644 --- a/modules/local/gtdbtk_db_preparation.nf +++ b/modules/local/gtdbtk_db_preparation.nf @@ -3,8 +3,8 @@ process GTDBTK_DB_PREPARATION { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path(database) diff --git a/modules/local/kraken2_db_preparation.nf b/modules/local/kraken2_db_preparation.nf index 81c294a8..38202110 100644 --- a/modules/local/kraken2_db_preparation.nf +++ b/modules/local/kraken2_db_preparation.nf @@ -2,8 +2,8 @@ process KRAKEN2_DB_PREPARATION { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path db diff --git a/modules/local/pool_paired_reads.nf b/modules/local/pool_paired_reads.nf index ac7fcb68..27c5e0f3 100644 --- a/modules/local/pool_paired_reads.nf +++ b/modules/local/pool_paired_reads.nf @@ -3,8 +3,8 @@ process POOL_PAIRED_READS { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: tuple val(meta), path(reads1), path(reads2) diff --git a/modules/local/pool_single_reads.nf b/modules/local/pool_single_reads.nf index ac165082..cc633d4a 100644 --- a/modules/local/pool_single_reads.nf +++ b/modules/local/pool_single_reads.nf @@ -3,8 +3,8 @@ process POOL_SINGLE_READS { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: tuple val(meta), path(reads) diff --git a/modules/local/quast_bins_summary.nf b/modules/local/quast_bins_summary.nf index 933758fb..eae5ef78 100644 --- a/modules/local/quast_bins_summary.nf +++ b/modules/local/quast_bins_summary.nf @@ -2,8 +2,8 @@ process QUAST_BINS_SUMMARY { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path(summaries) diff --git a/modules/nf-core/modules/gunzip/main.nf b/modules/nf-core/modules/gunzip/main.nf index 9d4b0666..61bf1afa 100644 --- a/modules/nf-core/modules/gunzip/main.nf +++ b/modules/nf-core/modules/gunzip/main.nf @@ -4,8 +4,8 @@ process GUNZIP { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: tuple val(meta), path(archive) From c1a30cdbb6202feda7d242c9b4795f05cd9b0971 Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Fri, 3 Jun 2022 08:47:53 +0200 Subject: [PATCH 221/246] Update CHANGELOG.md --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index c0c1b275..a147f8ab 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -19,6 +19,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint - [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths` for `[assembler]-[sample/group]-depth.txt.gz`, `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` - [#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` +- Replace base container for standard shell tools to fix problems with running on Google Cloud ### `Fixed` From 152c1b0d5430863c86fb7e0758846514548f5ffa Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Jun 2022 08:58:50 +0200 Subject: [PATCH 222/246] Add missing file --- modules/local/cat_db.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/cat_db.nf b/modules/local/cat_db.nf index 3642c939..03f463c9 100644 --- a/modules/local/cat_db.nf +++ b/modules/local/cat_db.nf @@ -3,8 +3,8 @@ process CAT_DB { conda (params.enable_conda ? "conda-forge::sed=4.7" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://containers.biocontainers.pro/s3/SingImgsRepo/biocontainers/v1.2.0_cv1/biocontainers_v1.2.0_cv1.img' : - 'biocontainers/biocontainers:v1.2.0_cv1' }" + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'ubuntu:20.04' }" input: path(database) From 6e760e3d8617baf0a9ed68567795619abac2ee68 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 10:08:34 +0200 Subject: [PATCH 223/246] Fix QUAST_BINS output description in output.md --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index fb5d2032..f8e5c484 100644 --- a/docs/output.md +++ b/docs/output.md @@ -381,9 +381,9 @@ For each genome bin the median sequencing depth is computed based on the corresp - `icarus_viewers/contig_size_viewer.html`: Diagram of contigs that are ordered from longest to shortest - `basic_stats/cumulative_plot.pdf`: Shows the growth of contig lengths (contigs are ordered from largest to shortest) - `basic_stats/GC_content_plot.pdf`: Shows the distribution of GC content in the contigs - - `basic_stats/[assembler]-[sample/group]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs + - `basic_stats/[assembler]-[bin]_GC_content_plot.pdf`: Histogram of the GC percentage for the contigs - `basic_stats/Nx_plot.pdf`: Plot of Nx values as x varies from 0 to 100%. - - `predicted_genes/[assembler]-[sample/group].rna.gff`: Contig positions for rRNA genes in gff version 3 format + - `predicted_genes/[assembler]-[bin].rna.gff`: Contig positions for rRNA genes in gff version 3 format - `predicted_genes/barrnap.log`: Barrnap log file (ribosomal RNA predictor) - `GenomeBinning/QC/` - `quast_summary.tsv`: QUAST output for all bins summarized From 24532274ee72c37dc728d0e977c4ccbc3b511399 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 10:10:43 +0200 Subject: [PATCH 224/246] Fix: use 'binner' info for GTDBTK and CAT --- conf/modules.config | 4 ++-- docs/output.md | 34 ++++++++++++++++---------------- modules/local/cat.nf | 20 +++++++++---------- modules/local/gtdbtk_classify.nf | 30 ++++++++++++++-------------- 4 files changed, 44 insertions(+), 44 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 1eafc2fe..f3cb1c42 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -277,7 +277,7 @@ process { withName: CAT { publishDir = [ - path: { "${params.outdir}/Taxonomy/CAT/${meta.assembler}" }, + path: { "${params.outdir}/Taxonomy/CAT/${meta.assembler}/${meta.binner}" }, mode: params.publish_dir_mode, saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] @@ -286,7 +286,7 @@ process { withName: GTDBTK_CLASSIFY { ext.args = "--extension fa" publishDir = [ - path: { "${params.outdir}/Taxonomy/GTDB-Tk/${meta.assembler}/${meta.id}" }, + path: { "${params.outdir}/Taxonomy/GTDB-Tk/${meta.assembler}/${meta.binner}/${meta.id}" }, mode: params.publish_dir_mode, pattern: "*.{log,tasv,tree.gz,fasta,fasta.gz}" ] diff --git a/docs/output.md b/docs/output.md index f8e5c484..89ab3ef4 100644 --- a/docs/output.md +++ b/docs/output.md @@ -437,15 +437,15 @@ Besides the reference files or output files created by BUSCO, the following summ

Output files -- `Taxonomy/CAT/[assembler]/` - - `[assembler]-[sample/group].ORF2LCA.names.txt.gz`: Tab-delimited files containing the lineage of each contig, with full lineage names - - `[assembler]-[sample/group].bin2classification.names.txt.gz`: Taxonomy classification of the genome bins, with full lineage names -- `Taxonomy/CAT/[assembler]/raw/` - - `[assembler]-[sample/group].concatenated.predicted_proteins.faa.gz`: Predicted protein sequences for each genome bin, in fasta format - - `[assembler]-[sample/group].concatenated.predicted_proteins.gff.gz`: Predicted protein features for each genome bin, in gff format - - `[assembler]-[sample/group].ORF2LCA.txt.gz`: Tab-delimited files containing the lineage of each contig - - `[assembler]-[sample/group].bin2classification.txt.gz`: Taxonomy classification of the genome bins - - `[assembler]-[sample/group].log`: Log files +- `Taxonomy/CAT/[assembler]/[binner]/` + - `[assembler]-[binner]-[sample/group].ORF2LCA.names.txt.gz`: Tab-delimited files containing the lineage of each contig, with full lineage names + - `[assembler]-[binner]-[sample/group].bin2classification.names.txt.gz`: Taxonomy classification of the genome bins, with full lineage names +- `Taxonomy/CAT/[assembler]/[binner]/raw/` + - `[assembler]-[binner]-[sample/group].concatenated.predicted_proteins.faa.gz`: Predicted protein sequences for each genome bin, in fasta format + - `[assembler]-[binner]-[sample/group].concatenated.predicted_proteins.gff.gz`: Predicted protein features for each genome bin, in gff format + - `[assembler]-[binner]-[sample/group].ORF2LCA.txt.gz`: Tab-delimited files containing the lineage of each contig + - `[assembler]-[binner]-[sample/group].bin2classification.txt.gz`: Taxonomy classification of the genome bins + - `[assembler]-[binner]-[sample/group].log`: Log files
@@ -465,14 +465,14 @@ If the parameters `--cat_db_generate` and `--save_cat_db` are set, additionally
Output files -- `Taxonomy/GTDB-Tk/[assembler]/[sample/group]/` - - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.summary.tsv`: Classifications for bacterial and archaeal genomes (see the [GTDB-Tk documentation for details](https://ecogenomics.github.io/GTDBTk/files/summary.tsv.html). - - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.classify.tree.gz`: Reference tree in Newick format containing query genomes placed with pplacer. - - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.markers_summary.tsv`: A summary of unique, duplicated, and missing markers within the 120 bacterial marker set, or the 122 archaeal marker set for each submitted genome. - - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.msa.fasta.gz`: FASTA file containing MSA of submitted and reference genomes. - - `gtdbtk.[assembler]-[sample/group].{bac120/ar122}.filtered.tsv`: A list of genomes with an insufficient number of amino acids in MSA. - - `gtdbtk.[assembler]-[sample/group].*.log`: Log files. - - `gtdbtk.[assembler]-[sample/group].failed_genomes.tsv`: A list of genomes for which the GTDB-Tk analysis failed, e.g. because Prodigal could not detect any genes. +- `Taxonomy/GTDB-Tk/[assembler]/[binner]/[sample/group]/` + - `gtdbtk.[assembler]-[binner]-[sample/group].{bac120/ar122}.summary.tsv`: Classifications for bacterial and archaeal genomes (see the [GTDB-Tk documentation for details](https://ecogenomics.github.io/GTDBTk/files/summary.tsv.html). + - `gtdbtk.[assembler]-[binner]-[sample/group].{bac120/ar122}.classify.tree.gz`: Reference tree in Newick format containing query genomes placed with pplacer. + - `gtdbtk.[assembler]-[binner]-[sample/group].{bac120/ar122}.markers_summary.tsv`: A summary of unique, duplicated, and missing markers within the 120 bacterial marker set, or the 122 archaeal marker set for each submitted genome. + - `gtdbtk.[assembler]-[binner]-[sample/group].{bac120/ar122}.msa.fasta.gz`: FASTA file containing MSA of submitted and reference genomes. + - `gtdbtk.[assembler]-[binner]-[sample/group].{bac120/ar122}.filtered.tsv`: A list of genomes with an insufficient number of amino acids in MSA. + - `gtdbtk.[assembler]-[binner]-[sample/group].*.log`: Log files. + - `gtdbtk.[assembler]-[binner]-[sample/group].failed_genomes.tsv`: A list of genomes for which the GTDB-Tk analysis failed, e.g. because Prodigal could not detect any genes. - `Taxonomy/GTDB-Tk/gtdbtk_summary.tsv`: A summary table of the GTDB-Tk classification results for all bins, also containing bins which were discarded based on the BUSCO QC, which were filtered out by GTDB-Tk ((listed in `*.filtered.tsv`) or for which the analysis failed (listed in `*.failed_genomes.tsv`).
diff --git a/modules/local/cat.nf b/modules/local/cat.nf index 009a4785..a4943c28 100644 --- a/modules/local/cat.nf +++ b/modules/local/cat.nf @@ -1,5 +1,5 @@ process CAT { - tag "${meta.assembler}-${meta.id}-${db_name}" + tag "${meta.assembler}-${meta.binner}-${meta.id}-${db_name}" conda (params.enable_conda ? "bioconda::cat=4.6 bioconda::diamond=2.0.6" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -21,18 +21,18 @@ process CAT { script: """ - CAT bins -b "bins/" -d database/ -t taxonomy/ -n "${task.cpus}" -s .fa --top 6 -o "${meta.assembler}-${meta.id}" --I_know_what_Im_doing - CAT add_names -i "${meta.assembler}-${meta.id}.ORF2LCA.txt" -o "${meta.assembler}-${meta.id}.ORF2LCA.names.txt" -t taxonomy/ - CAT add_names -i "${meta.assembler}-${meta.id}.bin2classification.txt" -o "${meta.assembler}-${meta.id}.bin2classification.names.txt" -t taxonomy/ + CAT bins -b "bins/" -d database/ -t taxonomy/ -n "${task.cpus}" -s .fa --top 6 -o "${meta.assembler}-${meta.binner}-${meta.id}" --I_know_what_Im_doing + CAT add_names -i "${meta.assembler}-${meta.binner}-${meta.id}.ORF2LCA.txt" -o "${meta.assembler}-${meta.binner}-${meta.id}.ORF2LCA.names.txt" -t taxonomy/ + CAT add_names -i "${meta.assembler}-${meta.binner}-${meta.id}.bin2classification.txt" -o "${meta.assembler}-${meta.binner}-${meta.id}.bin2classification.names.txt" -t taxonomy/ mkdir raw mv *.ORF2LCA.txt *.predicted_proteins.faa *.predicted_proteins.gff *.log *.bin2classification.txt raw/ - gzip "raw/${meta.assembler}-${meta.id}.ORF2LCA.txt" \ - "raw/${meta.assembler}-${meta.id}.concatenated.predicted_proteins.faa" \ - "raw/${meta.assembler}-${meta.id}.concatenated.predicted_proteins.gff" \ - "raw/${meta.assembler}-${meta.id}.bin2classification.txt" \ - "${meta.assembler}-${meta.id}.ORF2LCA.names.txt" \ - "${meta.assembler}-${meta.id}.bin2classification.names.txt" + gzip "raw/${meta.assembler}-${meta.binner}-${meta.id}.ORF2LCA.txt" \ + "raw/${meta.assembler}-${meta.binner}-${meta.id}.concatenated.predicted_proteins.faa" \ + "raw/${meta.assembler}-${meta.binner}-${meta.id}.concatenated.predicted_proteins.gff" \ + "raw/${meta.assembler}-${meta.binner}-${meta.id}.bin2classification.txt" \ + "${meta.assembler}-${meta.binner}-${meta.id}.ORF2LCA.names.txt" \ + "${meta.assembler}-${meta.binner}-${meta.id}.bin2classification.names.txt" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/gtdbtk_classify.nf b/modules/local/gtdbtk_classify.nf index a134d8be..077849b8 100644 --- a/modules/local/gtdbtk_classify.nf +++ b/modules/local/gtdbtk_classify.nf @@ -1,5 +1,5 @@ process GTDBTK_CLASSIFY { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" conda (params.enable_conda ? "bioconda::gtdbtk=1.5.0" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -11,16 +11,16 @@ process GTDBTK_CLASSIFY { tuple val(db_name), path("database/*") output: - path "gtdbtk.${meta.assembler}-${meta.id}.*.summary.tsv" , emit: summary - path "gtdbtk.${meta.assembler}-${meta.id}.*.classify.tree.gz" , emit: tree - path "gtdbtk.${meta.assembler}-${meta.id}.*.markers_summary.tsv", emit: markers - path "gtdbtk.${meta.assembler}-${meta.id}.*.msa.fasta.gz" , emit: msa - path "gtdbtk.${meta.assembler}-${meta.id}.*.user_msa.fasta" , emit: user_msa - path "gtdbtk.${meta.assembler}-${meta.id}.*.filtered.tsv" , emit: filtered - path "gtdbtk.${meta.assembler}-${meta.id}.log" , emit: log - path "gtdbtk.${meta.assembler}-${meta.id}.warnings.log" , emit: warnings - path "gtdbtk.${meta.assembler}-${meta.id}.failed_genomes.tsv" , emit: failed - path "versions.yml" , emit: versions + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.summary.tsv" , emit: summary + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.classify.tree.gz" , emit: tree + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.markers_summary.tsv", emit: markers + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.msa.fasta.gz" , emit: msa + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.user_msa.fasta" , emit: user_msa + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.*.filtered.tsv" , emit: filtered + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.log" , emit: log + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.warnings.log" , emit: warnings + path "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.failed_genomes.tsv" , emit: failed + path "versions.yml" , emit: versions script: def args = task.ext.args ?: '' @@ -33,7 +33,7 @@ process GTDBTK_CLASSIFY { gtdbtk classify_wf $args \ --genome_dir bins \ - --prefix "gtdbtk.${meta.assembler}-${meta.id}" \ + --prefix "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}" \ --out_dir "\${PWD}" \ --cpus ${task.cpus} \ --pplacer_cpus ${params.gtdbtk_pplacer_cpus} \ @@ -41,9 +41,9 @@ process GTDBTK_CLASSIFY { --min_perc_aa ${params.gtdbtk_min_perc_aa} \ --min_af ${params.gtdbtk_min_af} - gzip "gtdbtk.${meta.assembler}-${meta.id}".*.classify.tree "gtdbtk.${meta.assembler}-${meta.id}".*.msa.fasta - mv gtdbtk.log "gtdbtk.${meta.assembler}-${meta.id}.log" - mv gtdbtk.warnings.log "gtdbtk.${meta.assembler}-${meta.id}.warnings.log" + gzip "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}".*.classify.tree "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}".*.msa.fasta + mv gtdbtk.log "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.log" + mv gtdbtk.warnings.log "gtdbtk.${meta.assembler}-${meta.binner}-${meta.id}.warnings.log" cat <<-END_VERSIONS > versions.yml "${task.process}": From c666942dedebc4ec16ffd6a776d5b6466f7ec15c Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Fri, 3 Jun 2022 11:34:56 +0200 Subject: [PATCH 225/246] Update CHANGELOG.md Co-authored-by: Sabrina Krakau --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index a147f8ab..17650e93 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -19,7 +19,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint - [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths` for `[assembler]-[sample/group]-depth.txt.gz`, `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` - [#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` -- Replace base container for standard shell tools to fix problems with running on Google Cloud +- [#315](https://github.com/nf-core/mag/pull/315) - Replace base container for standard shell tools to fix problems with running on Google Cloud ### `Fixed` From c530a726fc4cdf4965ccb6cdacffec2cd2ad55fc Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Jun 2022 13:04:16 +0200 Subject: [PATCH 226/246] Activate institutional configs --- nextflow.config | 10 +++++----- 1 file changed, 5 insertions(+), 5 deletions(-) diff --git a/nextflow.config b/nextflow.config index 789673e2..43926a05 100644 --- a/nextflow.config +++ b/nextflow.config @@ -155,11 +155,11 @@ try { // Load nf-core/mag custom profiles from different institutions. // Warning: Uncomment only if a pipeline-specific instititutional config already exists on nf-core/configs! -// try { -// includeConfig "${params.custom_config_base}/pipeline/mag.config" -// } catch (Exception e) { -// System.err.println("WARNING: Could not load nf-core/config/mag profiles: ${params.custom_config_base}/pipeline/mag.config") -// } +try { + includeConfig "${params.custom_config_base}/pipeline/mag.config" +} catch (Exception e) { + System.err.println("WARNING: Could not load nf-core/config/mag profiles: ${params.custom_config_base}/pipeline/mag.config") +} profiles { From 17c496c1bbec0d599f72144de78e27415d13a03f Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Jun 2022 13:07:28 +0200 Subject: [PATCH 227/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index c69ceeba..0b381d2d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#263](https://github.com/nf-core/mag/pull/263) - Add MaxBin2 as second contig binning tool - [#285](https://github.com/nf-core/mag/pull/285) - Add AdapterRemoval2 as an alternative read trimmer - [#291](https://github.com/nf-core/mag/pull/291) - Add DAS Tool for bin refinement +- [#319](https://github.com/nf-core/mag/pull/319) - Activate pipeline-specific institutional nf-core/configs ### `Changed` From 24efccf5224def79cc1dfd1a58e018a78e3d25d8 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 14:24:15 +0200 Subject: [PATCH 228/246] Update modules/local/bowtie2_assembly_align.nf Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- modules/local/bowtie2_assembly_align.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/modules/local/bowtie2_assembly_align.nf b/modules/local/bowtie2_assembly_align.nf index 6402962a..9cac97d0 100644 --- a/modules/local/bowtie2_assembly_align.nf +++ b/modules/local/bowtie2_assembly_align.nf @@ -26,7 +26,6 @@ process BOWTIE2_ASSEMBLY_ALIGN { $args \\ $input \\ 2> "${name}.bowtie2.log" | \ - samtools view -@ "${task.cpus}" -bS | \ samtools sort -@ "${task.cpus}" -o "${name}.bam" samtools index "${name}.bam" From f1886d6449b2a8641c64dda69cbdde71f68ef1a5 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 14:33:49 +0200 Subject: [PATCH 229/246] Update CHANGELOG.md Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- CHANGELOG.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c69ceeba..e11524e2 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -15,10 +15,9 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Changed` -- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289) - Update to nf-core 2.3.2 `TEMPLATE` +- [#269](https://github.com/nf-core/mag/pull/269),[#283](https://github.com/nf-core/mag/pull/283),[#289](https://github.com/nf-core/mag/pull/289),[#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` - [#286](https://github.com/nf-core/mag/pull/286) - Cite our publication instead of the preprint - [#291](https://github.com/nf-core/mag/pull/291), [#299](https://github.com/nf-core/mag/pull/299) - Add extra results folder `GenomeBinning/depths/contigs` for `[assembler]-[sample/group]-depth.txt.gz`, and `GenomeBinning/depths/bins` for `bin_depths_summary.tsv` and `[assembler]-[binner]-[sample/group]-binDepths.heatmap.png` -- [#302](https://github.com/nf-core/mag/pull/302) - Update to nf-core 2.4 `TEMPLATE` - [#315](https://github.com/nf-core/mag/pull/315) - Replace base container for standard shell tools to fix problems with running on Google Cloud ### `Fixed` From cad0db3e09973429a56bc50256d54dd9ee9ffc1f Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 14:42:16 +0200 Subject: [PATCH 230/246] Update docs/output.md Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 89ab3ef4..f28e3920 100644 --- a/docs/output.md +++ b/docs/output.md @@ -353,7 +353,7 @@ By default, only the raw bins (and unbinned contigs) from the actual binning met ### Bin sequencing depth -For each genome bin the median sequencing depth is computed based on the corresponding contig depths of each bin or refined bin. +For each bin or refined bin the median sequencing depth is computed based on the corresponding contig depths.
Output files From 053f811b7207c6b3a0d967ddc6375e3d89723e27 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 15:08:43 +0200 Subject: [PATCH 231/246] Apply suggestions from code review Co-authored-by: James A. Fellows Yates --- CHANGELOG.md | 7 +------ CITATIONS.md | 5 +++-- docs/output.md | 6 +++--- docs/usage.md | 2 +- modules/local/rename_predastool.nf | 2 +- subworkflows/local/binning.nf | 5 +++-- subworkflows/local/binning_refinement.nf | 2 +- 7 files changed, 13 insertions(+), 16 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e11524e2..24bd3537 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v2.2.0 - 2022/06/02 +## v2.2.0 - 2022/06/03 ### `Added` @@ -29,11 +29,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Dependencies` -| Tool | Previous version | New version | -| -------- | ---------------- | ----------- | -| bcftools | 1.13 | 1.14 | -| samtools | 1.14 | 1.15 | -| pydamage | 0.62 | 0.70 | ## v2.1.1 - 2021/11/25 diff --git a/CITATIONS.md b/CITATIONS.md index 33337945..a46ab6a8 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -96,6 +96,9 @@ > Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014 Jul 15;30(14):2068-9. doi: 10.1093/bioinformatics/btu153. Epub 2014 Mar 18. PMID: 24642063. +- [PyDamage](https://doi.org/10.7717/peerj.11845) + > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 + - [SAMtools](https://doi.org/10.1093/bioinformatics/btp352) > Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., … 1000 Genome Project Data Processing Subgroup. (2009). The Sequence Alignment/Map format and SAMtools. Bioinformatics , 25(16), 2078–2079. doi: 10.1093/bioinformatics/btp352. @@ -104,8 +107,6 @@ > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. -- [PyDamage](https://doi.org/10.7717/peerj.11845) - > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 ## Data diff --git a/docs/output.md b/docs/output.md index f28e3920..7d00380b 100644 --- a/docs/output.md +++ b/docs/output.md @@ -256,7 +256,7 @@ Protein-coding genes are predicted for each assembly. ### Contig sequencing depth -Sequencing depth per contig and sample is generated by `jgi_summarize_bam_contig_depths --outputDepth`. The values correspond to `(sum of exactly aligned bases) / ((contig length)-2*75)`. For example, for two reads aligned exactly with `10` and `9` bases on a 1000 bp long contig the depth is calculated by `(10+9)/(1000-2*75)` (1000bp length of contig minus 75bp from each end, which is excluded). +Sequencing depth per contig and sample is generated by MetaBAT2's `jgi_summarize_bam_contig_depths --outputDepth`. The values correspond to `(sum of exactly aligned bases) / ((contig length)-2*75)`. For example, for two reads aligned exactly with `10` and `9` bases on a 1000 bp long contig the depth is calculated by `(10+9)/(1000-2*75)` (1000bp length of contig minus 75bp from each end, which is excluded). These depth files are used for downstream binning steps. @@ -349,7 +349,7 @@ Files in these two folders contain all contigs of an assembly. By default, only the raw bins (and unbinned contigs) from the actual binning methods, but not from the binning refinement with DAS Tool, will be used for downstream bin quality control, annotation and taxonomic classification. The parameter `--postbinning_input` can be used to change this behaviour. -⚠️ Due to ability to perform downstream QC of both raw and refined bins in parallel (via `--postbinning_input)`, bin names in DAS Tools's `*_allBins.eval` file will include `Refined`, however for this particular file, they _actually_ refer to the 'raw' input bins. The pipeline renames the input files prior to running DASTool to ensure they can be disambuguated from the original bin files in the downstream QC steps. +⚠️ Due to ability to perform downstream QC of both raw and refined bins in parallel (via `--postbinning_input)`, bin names in DAS Tools's `*_allBins.eval` file will include `Refined`. However for this particular file, they _actually_ refer to the 'raw' input bins. The pipeline renames the input files prior to running DASTool to ensure they can be disambuguated from the original bin files in the downstream QC steps. ### Bin sequencing depth @@ -515,7 +515,7 @@ Whole genome annotation is the process of identifying features of interest in a ### `PyDamage` -[Pydamage](https://github.com/maxibor/pydamage), is a tool to automate the process of ancient DNA damage identification and estimation +[Pydamage](https://github.com/maxibor/pydamage), is a tool to automate the process of ancient DNA damage identification and estimation from contigs. After modelling the ancient DNA damage using the C to T transitions, Pydamage uses a likelihood ratio test to discriminate between truly ancient, and modern contigs originating from sample contamination.
Output files diff --git a/docs/usage.md b/docs/usage.md index b24e000f..ca9d54a4 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -286,7 +286,7 @@ NXF_OPTS='-Xms1g -Xmx4g' MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: -[Characteristic patterns of ancient DNA (aDNA) damage](<(https://doi.org/10.1073/pnas.0704665104)>), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), MAG can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. +[Characteristic patterns of ancient DNA (aDNA) damage](<(https://doi.org/10.1073/pnas.0704665104)>), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), nf-core/mag can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. ## A note on bin refinement diff --git a/modules/local/rename_predastool.nf b/modules/local/rename_predastool.nf index af670f4e..2250de79 100644 --- a/modules/local/rename_predastool.nf +++ b/modules/local/rename_predastool.nf @@ -1,5 +1,5 @@ process RENAME_PREDASTOOL { - tag "${meta.assembler}-${meta.id}" + tag "${meta.assembler}-${meta.binner}-${meta.id}" label 'process_low' // Using container from multiqc since it'll be included anyway diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 9f19e2cd..67243638 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -126,8 +126,9 @@ workflow BINNING { // Compute bin depths for different samples (according to `binning_map_mode`) // Have to remove binner meta before joining with according depths files, - // as required for MAG_DEPTHS - // add 'binner' info again and finally group by 'assembler', 'id', 'binner' + // as required for MAG_DEPTHS, but we can add 'binner' + // info again based on file name and finally group by + // 'assembler', 'id', 'binner' ch_depth_input = ch_binning_results_gunzipped .mix(ch_splitfasta_results_gunzipped ) .map { meta, bin -> diff --git a/subworkflows/local/binning_refinement.nf b/subworkflows/local/binning_refinement.nf index 17276583..84e0e142 100644 --- a/subworkflows/local/binning_refinement.nf +++ b/subworkflows/local/binning_refinement.nf @@ -21,7 +21,7 @@ def getColNo(filename) { workflow BINNING_REFINEMENT { take: - contigs // + contigs bins // channel: [ val(meta), path(bins) ] depths reads From f30a3229f6dc5119de9e53fea7b976436ff94e41 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Fri, 3 Jun 2022 21:30:52 +0200 Subject: [PATCH 232/246] Apply prettier --- CHANGELOG.md | 1 - CITATIONS.md | 2 +- 2 files changed, 1 insertion(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 24bd3537..e483bc2d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -29,7 +29,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Dependencies` - ## v2.1.1 - 2021/11/25 ### `Added` diff --git a/CITATIONS.md b/CITATIONS.md index a46ab6a8..cef607f0 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -97,6 +97,7 @@ > Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014 Jul 15;30(14):2068-9. doi: 10.1093/bioinformatics/btu153. Epub 2014 Mar 18. PMID: 24642063. - [PyDamage](https://doi.org/10.7717/peerj.11845) + > Borry M, Hübner A, Rohrlach AB, Warinner C. 2021. PyDamage: automated ancient damage identification and estimation for contigs in ancient DNA de novo assembly. PeerJ 9:e11845 doi: 10.7717/peerj.11845 - [SAMtools](https://doi.org/10.1093/bioinformatics/btp352) @@ -107,7 +108,6 @@ > Nurk, S., Meleshko, D., Korobeynikov, A., & Pevzner, P. A. (2017). metaSPAdes: a new versatile metagenomic assembler. Genome research, 27(5), 824-834. doi: 10.1101/gr.213959.116. - ## Data - [Full-size test data](https://doi.org/10.1038/s41587-019-0191-2) From a2f5fe9a83982ed44c289200f58a5e82e31340b7 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 11:48:44 +0200 Subject: [PATCH 233/246] Update subworkflows/local/binning.nf Co-authored-by: James A. Fellows Yates --- subworkflows/local/binning.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/subworkflows/local/binning.nf b/subworkflows/local/binning.nf index 67243638..0c7f4c28 100644 --- a/subworkflows/local/binning.nf +++ b/subworkflows/local/binning.nf @@ -169,8 +169,6 @@ workflow BINNING { ch_binning_results_gunzipped_final = ch_binning_results_gunzipped.groupTuple(by: 0) ch_binning_results_gzipped_final = ch_binning_results_gzipped_final.groupTuple(by: 0) - SPLIT_FASTA.out.unbinned - emit: bins = ch_binning_results_gunzipped_final bins_gz = ch_binning_results_gzipped_final From e20999186ca2a715af6fcc725aa256e10850a9b1 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Fri, 3 Jun 2022 14:41:48 +0200 Subject: [PATCH 234/246] Add --outdir to usage describtion in test configs --- conf/test_adapterremoval.config | 2 +- conf/test_ancient_dna.config | 2 +- conf/test_busco_auto.config | 2 +- conf/test_host_rm.config | 2 +- conf/test_hybrid.config | 2 +- conf/test_hybrid_host_rm.config | 2 +- 6 files changed, 6 insertions(+), 6 deletions(-) diff --git a/conf/test_adapterremoval.config b/conf/test_adapterremoval.config index 0725cbc2..45836fbc 100644 --- a/conf/test_adapterremoval.config +++ b/conf/test_adapterremoval.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test, + nextflow run nf-core/mag -profile test, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_ancient_dna.config b/conf/test_ancient_dna.config index 84e18830..33b6f4f8 100644 --- a/conf/test_ancient_dna.config +++ b/conf/test_ancient_dna.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test_ancient_dna, + nextflow run nf-core/mag -profile test_ancient_dna, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_busco_auto.config b/conf/test_busco_auto.config index 8e3d3f86..adf3d277 100644 --- a/conf/test_busco_auto.config +++ b/conf/test_busco_auto.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test_busco_auto, + nextflow run nf-core/mag -profile test_busco_auto, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_host_rm.config b/conf/test_host_rm.config index 4e912bd3..c93317db 100644 --- a/conf/test_host_rm.config +++ b/conf/test_host_rm.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test_host_rm, + nextflow run nf-core/mag -profile test_host_rm, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_hybrid.config b/conf/test_hybrid.config index edc879e7..873d2c5c 100644 --- a/conf/test_hybrid.config +++ b/conf/test_hybrid.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test_hybrid, + nextflow run nf-core/mag -profile test_hybrid, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_hybrid_host_rm.config b/conf/test_hybrid_host_rm.config index 08649bc5..29db0f7a 100644 --- a/conf/test_hybrid_host_rm.config +++ b/conf/test_hybrid_host_rm.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/mag -profile test_hybrid_host_rm, + nextflow run nf-core/mag -profile test_hybrid_host_rm, --outdir ---------------------------------------------------------------------------------------- */ From 023873eaf853fd0765cf31a9bea1712b59d51bd4 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 15:45:33 +0200 Subject: [PATCH 235/246] Change remaining ext.suffix to ext.prefix --- conf/modules.config | 8 ++++---- modules/local/bowtie2_removal_align.nf | 4 ++-- modules/local/nanoplot.nf | 4 ++-- 3 files changed, 8 insertions(+), 8 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index f3cb1c42..846c518c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -75,7 +75,7 @@ process { } withName: BOWTIE2_PHIX_REMOVAL_ALIGN { - ext.suffix = "phix_removed" + ext.prefix = { "${meta.id}.phix_removed" } publishDir = [ path: { "${params.outdir}/QC_shortreads/remove_phix" }, mode: params.publish_dir_mode, @@ -84,7 +84,7 @@ process { } withName: BOWTIE2_HOST_REMOVAL_ALIGN { - ext.suffix = "host_removed" + ext.prefix = { "${meta.id}.host_removed" } publishDir = [ path: { "${params.outdir}/QC_shortreads/remove_host" }, mode: params.publish_dir_mode, @@ -111,7 +111,7 @@ process { } withName: NANOPLOT_RAW { - ext.suffix = 'raw' + ext.prefix = 'raw' publishDir = [ path: { "${params.outdir}/QC_longreads/NanoPlot/${meta.id}" }, mode: params.publish_dir_mode, @@ -120,7 +120,7 @@ process { } withName: NANOPLOT_FILTERED { - ext.suffix = 'filtered' + ext.prefix = 'filtered' publishDir = [ path: { "${params.outdir}/QC_longreads/NanoPlot/${meta.id}" }, mode: params.publish_dir_mode, diff --git a/modules/local/bowtie2_removal_align.nf b/modules/local/bowtie2_removal_align.nf index 5d11e916..880d932f 100644 --- a/modules/local/bowtie2_removal_align.nf +++ b/modules/local/bowtie2_removal_align.nf @@ -2,7 +2,7 @@ * Bowtie2 for read removal */ process BOWTIE2_REMOVAL_ALIGN { - tag "${meta.id}-${task.ext.suffix}" + tag "$meta.id" conda (params.enable_conda ? "bioconda::bowtie2=2.4.2" : null) container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -21,7 +21,7 @@ process BOWTIE2_REMOVAL_ALIGN { script: def args = task.ext.args ?: '' - def prefix = task.ext.suffix ? "${meta.id}.${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ?: "${meta.id}" def sensitivity = params.host_removal_verysensitive ? "--very-sensitive" : "--sensitive" def save_ids = params.host_removal_save_ids ? "Y" : "N" if (!meta.single_end){ diff --git a/modules/local/nanoplot.nf b/modules/local/nanoplot.nf index f280aef5..f746e8c5 100644 --- a/modules/local/nanoplot.nf +++ b/modules/local/nanoplot.nf @@ -16,8 +16,8 @@ process NANOPLOT { path "versions.yml" , emit: versions script: - def prefix = task.ext.suffix ? "-p ${task.ext.suffix}_" : '' - def title = task.ext.suffix ? "${meta.id}_${task.ext.suffix}" : "${meta.id}" + def prefix = task.ext.prefix ? "-p ${task.ext.prefix}_" : '' + def title = task.ext.prefix ? "${meta.id}_${task.ext.prefix}" : "${meta.id}" """ NanoPlot -t ${task.cpus} \ ${prefix} \ From a36fe42cfc3289b5fbbde0108a206ed7b2135c63 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 17:41:43 +0200 Subject: [PATCH 236/246] Fix parameter processing in BOWTIE2_REMOVAL_ALIGN --- conf/modules.config | 2 ++ modules/local/bowtie2_removal_align.nf | 10 +++++----- 2 files changed, 7 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 846c518c..4051ec08 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -84,6 +84,8 @@ process { } withName: BOWTIE2_HOST_REMOVAL_ALIGN { + ext.args = params.host_removal_verysensitive ? "--very-sensitive" : "--sensitive" + ext.args2 = params.host_removal_save_ids ? "--host_removal_save_ids" : '' ext.prefix = { "${meta.id}.host_removed" } publishDir = [ path: { "${params.outdir}/QC_shortreads/remove_host" }, diff --git a/modules/local/bowtie2_removal_align.nf b/modules/local/bowtie2_removal_align.nf index 880d932f..919111c0 100644 --- a/modules/local/bowtie2_removal_align.nf +++ b/modules/local/bowtie2_removal_align.nf @@ -21,15 +21,15 @@ process BOWTIE2_REMOVAL_ALIGN { script: def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - def sensitivity = params.host_removal_verysensitive ? "--very-sensitive" : "--sensitive" - def save_ids = params.host_removal_save_ids ? "Y" : "N" + def args2 = task.ext.args2 ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def save_ids = (args2.contains('--host_removal_save_ids')) ? "Y" : "N" if (!meta.single_end){ """ bowtie2 -p ${task.cpus} \ -x ${index[0].getSimpleName()} \ -1 "${reads[0]}" -2 "${reads[1]}" \ - $sensitivity \ + $args \ --un-conc-gz ${prefix}.unmapped_%.fastq.gz \ --al-conc-gz ${prefix}.mapped_%.fastq.gz \ 1> /dev/null \ @@ -50,7 +50,7 @@ process BOWTIE2_REMOVAL_ALIGN { bowtie2 -p ${task.cpus} \ -x ${index[0].getSimpleName()} \ -U ${reads} \ - $sensitivity \ + $args \ --un-gz ${prefix}.unmapped.fastq.gz \ --al-gz ${prefix}.mapped.fastq.gz \ 1> /dev/null \ From 77ec9c4921d8282bb9d27397ac2cc7bcdd8fe0f4 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 17:44:18 +0200 Subject: [PATCH 237/246] Fix docs for host removal --- docs/output.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 7d00380b..52752511 100644 --- a/docs/output.md +++ b/docs/output.md @@ -89,7 +89,8 @@ The pipeline uses bowtie2 to map short reads against the host reference genome s Output files - `QC_shortreads/remove_host/` - - `[sample].host_removed.bowtie2.log`: Contains the bowtie2 log file indicating how many reads have been mapped as well as a file listing the read ids of discarded reads. + - `[sample].host_removed.bowtie2.log`: Contains the bowtie2 log file indicating how many reads have been mapped. + - `[sample].host_removed.mapped*.read_ids.txt`: Contains a file listing the read ids of discarded reads.
From 945f3ef4982d3d3508b47afdba976e31b28babbc Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 17:47:32 +0200 Subject: [PATCH 238/246] Adjust TOC in output.md --- docs/output.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 52752511..1c366afe 100644 --- a/docs/output.md +++ b/docs/output.md @@ -18,6 +18,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d - [Taxonomic classification of binned genomes](#taxonomic-classification-of-binned-genomes) - [Genome annotation of binned genomes](#genome-annotation-of-binned-genomes) - [Additional summary for binned genomes](#additional-summary-for-binned-genomes) +- [Ancient DNA](#ancient-dna) - [MultiQC](#multiqc) - aggregate report, describing results of the whole pipeline - [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution @@ -512,7 +513,8 @@ Whole genome annotation is the process of identifying features of interest in a
-## Ancient DNA (optional, only with `-profile ancient_dna`) +## Ancient DNA +Optional, only running when parameter `-profile ancient_dna` is specified. ### `PyDamage` From 1930a219219201bc798b9683c82ceb4cebbfbbf7 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 22:45:49 +0200 Subject: [PATCH 239/246] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d7136e61..fed0e457 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -27,6 +27,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#305](https://github.com/nf-core/mag/pull/305) - Add missing Bowtie2 version for process `BOWTIE2_PHIX_REMOVAL_ALIGN` to `software_versions.yml` - [#307](https://github.com/nf-core/mag/pull/307) - Fix retrieval of GTDB-Tk version (note about newer version caused error in `CUSTOM_DUMPSOFTWAREVERSIONS`) - [#309](https://github.com/nf-core/mag/pull/309) - Fix publishing of BUSCO `busco_downloads/` folder, i.e. publish only when `--save_busco_reference` is specified +- [#321](https://github.com/nf-core/mag/pull/321) - Fix parameter processing in `BOWTIE2_REMOVAL_ALIGN` (which was erroneously for `BOWTIE2_PHIX_REMOVAL_ALIGN`) ### `Dependencies` From 5d909952b04d51fe054e5949f22afde6526c51de Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Wed, 8 Jun 2022 23:14:07 +0200 Subject: [PATCH 240/246] Add tool version changes to CHANGELOG --- CHANGELOG.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index fed0e457..c250fc70 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -31,6 +31,11 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Dependencies` +| Tool | Previous version | New version | +| -------- | ---------------- | ----------- | +| fastp | 0.20.1 | 0.23.2 | +| MULTIQC | 1.11 | 1.12 | + ## v2.1.1 - 2021/11/25 ### `Added` From 93a778c34aea9f116b06d539cbc1f088ea5840b9 Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Thu, 9 Jun 2022 08:31:47 +0200 Subject: [PATCH 241/246] Prettier --- CHANGELOG.md | 8 ++++---- docs/output.md | 1 + 2 files changed, 5 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c250fc70..84c25086 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -31,10 +31,10 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### `Dependencies` -| Tool | Previous version | New version | -| -------- | ---------------- | ----------- | -| fastp | 0.20.1 | 0.23.2 | -| MULTIQC | 1.11 | 1.12 | +| Tool | Previous version | New version | +| ------- | ---------------- | ----------- | +| fastp | 0.20.1 | 0.23.2 | +| MULTIQC | 1.11 | 1.12 | ## v2.1.1 - 2021/11/25 diff --git a/docs/output.md b/docs/output.md index 1c366afe..c969b9bf 100644 --- a/docs/output.md +++ b/docs/output.md @@ -514,6 +514,7 @@ Whole genome annotation is the process of identifying features of interest in a ## Ancient DNA + Optional, only running when parameter `-profile ancient_dna` is specified. ### `PyDamage` From c7d9edd407f5df09e0bab6c5981230d630ac5978 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 13 Jun 2022 11:45:56 +0200 Subject: [PATCH 242/246] Fix CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 84c25086..03f80ba2 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -34,7 +34,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 | Tool | Previous version | New version | | ------- | ---------------- | ----------- | | fastp | 0.20.1 | 0.23.2 | -| MULTIQC | 1.11 | 1.12 | +| MultiQC | 1.11 | 1.12 | ## v2.1.1 - 2021/11/25 From 9806efbedbad09d28e2932e37a0467f45896b089 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 13 Jun 2022 12:25:02 +0200 Subject: [PATCH 243/246] Update docs/output.md Co-authored-by: James A. Fellows Yates --- docs/output.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/docs/output.md b/docs/output.md index 7d00380b..081aa1fd 100644 --- a/docs/output.md +++ b/docs/output.md @@ -333,6 +333,8 @@ Files in these two folders contain all contigs of an assembly. [DAS Tool](https://github.com/cmks/DAS_Tool) is an automated binning refinement method that integrates the results of a flexible number of binning algorithms to calculate an optimized, non-redundant set of bins from a single assembly. nf-core/mag uses this tool to attempt to further improve bins based on combining the MetaBAT2 and MaxBin2 binning output, assuming sufficient quality is met for those bins. +DAS Tool will remove contigs from bins that do not pass additional filtering criteria, and will discard redundant lower-quality output from binners that represent the same estimated 'organism', until the single highest quality bin is represented. +
Output files From 7e5275cf456d8fae754b1c40967c40b5fe9baf1f Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 13 Jun 2022 12:30:47 +0200 Subject: [PATCH 244/246] Update docs/usage.md --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index ca9d54a4..ef7fccb3 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -284,7 +284,7 @@ NXF_OPTS='-Xms1g -Xmx4g' ## A note on the ancient DNA subworkflow -MAG integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: +nf-core/mag integrates an additional subworkflow to validate ancient DNA _de novo_ assembly: [Characteristic patterns of ancient DNA (aDNA) damage](<(https://doi.org/10.1073/pnas.0704665104)>), namely DNA fragmentation and cytosine deamination (observed as C-to-T transitions) are typically used to authenticate aDNA sequences. By identifying assembled contigs carrying typical aDNA damages using [PyDamage](https://github.com/maxibor/pydamage), nf-core/mag can report and distinguish ancient contigs from contigs carrying no aDNA damage. Furthermore, to mitigate the effect of aDNA damage on contig sequence assembly, [freebayes](https://github.com/freebayes/freebayes) in combination with [BCFtools](https://github.com/samtools/bcftools) are used to (re)call the variants from the reads aligned to the contigs, and (re)generate contig consensus sequences. From a7d90a79309e4ffd93de77ba68680757731474a1 Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 13 Jun 2022 14:46:22 +0200 Subject: [PATCH 245/246] Update authors in README --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 1d68e974..e666ce61 100644 --- a/README.md +++ b/README.md @@ -88,7 +88,7 @@ When group-wise co-assembly is enabled, `SPAdes` is run on accordingly pooled re ## Credits -nf-core/mag was written by [Hadrien Gourlé](https://hadriengourle.com) at [SLU](https://slu.se), [Daniel Straub](https://github.com/d4straub) and [Sabrina Krakau](https://github.com/skrakau) at the [Quantitative Biology Center (QBiC)](http://qbic.life). +nf-core/mag was written by [Hadrien Gourlé](https://hadriengourle.com) at [SLU](https://slu.se), [Daniel Straub](https://github.com/d4straub) and [Sabrina Krakau](https://github.com/skrakau) at the [Quantitative Biology Center (QBiC)](http://qbic.life). [James A. Fellows Yates](https://github.com/jfy133) and [Maxime Borry](https://github.com/maxibor) at the [Max Planck Institute for Evolutionary Anthropology](https://www.eva.mpg.de) joined in version 2.2.0. Long read processing was inspired by [caspargross/HybridAssembly](https://github.com/caspargross/HybridAssembly) written by Caspar Gross [@caspargross](https://github.com/caspargross) From 6cff6093946ce52871c700ba1bb956d5aa54c2ee Mon Sep 17 00:00:00 2001 From: Sabrina Krakau Date: Mon, 13 Jun 2022 14:48:32 +0200 Subject: [PATCH 246/246] Update CHANGELOG.md Co-authored-by: James A. Fellows Yates --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 03f80ba2..c25942bd 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v2.2.0 - 2022/06/03 +## v2.2.0 - 2022/06/14 ### `Added`

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+ + + nf-core/mag Pipeline Report + + +
+ - +

nf-core/mag v${version}

+

Run Name: $runName

-

nf-core/mag v${version}

-

Run Name: $runName

- -<% if (!success){ - out << """ -
-

nf-core/mag execution completed unsuccessfully!

+ <% if (!success){ out << """ +
+

nf-core/mag execution completed unsuccessfully!

The exit status of the task that caused the workflow execution to fail was: $exitStatus.

The full error message was:

- 
${errorReport}
-
- """ -} else { - out << """ -
+ 
${errorReport}
+
+ """ } else { out << """ +
nf-core/mag execution completed successfully! -
- """ -} -%> +
+ """ } %> -

The workflow was completed at $dateComplete (duration: $duration)

-

The command used to launch the workflow was as follows:

-
$commandLine
+

The workflow was completed at $dateComplete (duration: $duration)

+

The command used to launch the workflow was as follows:

+ 
+$commandLine
-

Pipeline Configuration:

- - - <% out << summary.collect{ k,v -> "" }.join("\n") %> - -
$k
$v
+

Pipeline Configuration:

+ + + <% out << summary.collect{ k,v -> " + + + + + " }.join("\n") %> + +
+ $k + + 
$v
+
-

nf-core/mag

-

https://github.com/nf-core/mag

- -
- - +

nf-core/mag

+

https://github.com/nf-core/mag

+