- Introduction
- Prerequisites
- Installation
- Setup
- Workflow Invocation
- Sweep Definition
- Implemented Workflows
- Included Tools
Meta-Sweeper is a tool for conducting parametric (combinatorial) sweeps of selected experimental variables relevant to metagenomics research. In particular, we have implemented workflows which enable the generation of simulated microbial communities and their simulated sequencing (WGS and HiC/3C). Downstream of simulation, specific topics of analysis in HiC/3C clustering and Non-negative matrix factorization have been provided. Built upon the Nextflow framework, Meta-Sweeper is general enough that there is no reason that alternative workflows or extensions to existing workflows could not be implemented.
Meta-Sweeper is the culmination and refinement of a central process employed in our ongoing work exploring how to obtain the maximum value when integrating HiC/3C sequencing data with conventional whole-genome shotgun DNA sequencing. To accomplish this, we conducted a parametric sweep of select experimental and model parameters such as sequencing depth, community composition and evolutionary divergence. At each sampled point in the sweep, we pitted a set of algorithms against each-other, where their aim was (in a sense) the deconvolution of the simulated community. This work culminated in the publication: Deconvoluting simulated metagenomes: the performance of hard- and soft- clustering algorithms applied to metagenomic chromosome conformation capture (3C).
Parametric sweeps, and the sampling of parameter spaces in general, have reoccurring applicability within bioinformatics beyond our own topic of research. We believe that this systematic approach is potentially useful to a broad range of research topics, offering a means for more thorough quality assurance and performance testing. Aside from explicit testing, the process of sampling a parameter space can also be used simply as an exploratory technique. In all cases, a reproducible approach is preferable to one-off ad-hoc approaches that can prove difficult to reproduce even for the original authors.
Some suggested applications are such things as: algorithm development, the exploration of experimental requirements for new techniques or simply the comparative assessment of existing tools.
Although numerous workflow management systems exist, we feel there is a lack of facility for parametric sweeps. Further, ease of deployment to various high-throughput computing environments is a crucial feature, when bioinformatics tasks are often computationally expensive in terms of CPU and/or memory resources.
To that end, we invested the time to resolve our processes into a more coherent and easily deployed system. We hope this will prove useful to others.
- Python 2.7
- Java SE 8+
- Method calls by Groovy and MetaSweeper.groovy
- Linux x86-64 runtime environment
- supplied external binary tools
- GCC
- if building Biopython (via pip)
- Python development library and headers
- Debian package: python-dev
- Redhat package: python-devel
- zlib library and headers
- Package for Debian: zlib1g-dev
- Package for Redhat: zlib-devel
- Beagle Library
- if using time-series workflow
- requires autoconf tool-chain.
- process well documented at: beagle-lib
The Nextflow framework, which can be installed easily using either of the following:
wget -qO- get.nextflow.io | bash or curl -fsSL get.nextflow.io | bash
This assumes you have met the prerequisite of installing Java 8+. Note, you will need to have wget or curl installed depending on your choice above. Whether either is installed by default is dependent on which distribution of Linux you are using.
In addition, meta-sweeper expects that the main executable nextflow is accessible on the path. Users can move this file to a location already on the path or add its parent directory to the path.
We are employing submodules, therefore when cloning the repository it is easiest to add the recurisve option.
git clone --recursive https://github.com/cerebis/meta-sweeper.gitThe workflows depend on the following Python modules, which must be installed prior to using it:
- biopython
- dendropy
- intervaltree
- networkx
- numpy
- pandas
- python-louvain
- pysam
- PyYaml
- scipy
- tqdm
- numexpr
Some of these dependencies can be installed using distributional package manager, but not all will be found.
sudo apt-get install python-biopython python-pandas python-yaml python-networkx python-pysamsudo yum install python-biopython python-pandas python-yaml python-networkx python-pysamA more general and complete solution is to use pip.
Using '--upgrade' will ensure that the current version of each module is installed. Though we have not encountered this problem, a note of caution. Upgrading Python modules through Pip, and thus outside of your system's package manager, does incur the potential risk of version conflicts if your installed system packages depend on obsolete or otherwise deprecated functionality with a given Python module. If in doubt, you could try the same command below but omit the '--upgrade' option.
pip install --upgrade -r requirements.txtBefore running a meta-sweeper workflow, you must initialise the shell environment.
- Set meta-sweepers home to your installed location.
- If meta-sweeper was installed in /home/user/meta-sweeper.
export METASWEEPER_HOME=/home/user/meta-sweeper
- Append the meta-sweeper home patht to nextflow's classpath.
export NXF_CLASSPATH=$NXF_CLASSPATH:$METASWEEPER_HOME
- Check that nextflow is installed and has been added to the path.
- Either of the following should return the canonical path to the nextflow executable.
command -v nextfloworwhich nextflow
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A further dependency on beagle-lib exists for the timeseries workflow.
To use this workflow, users must set the environmental variable BEAGLE_LIBDIR to point to the directory containing
libhmsbeagle-jni.soshared library file.E.g.
BEAGLE_LIBDIR=/usr/libFor some Linux distributions, beagle-lib can be satisfied through the system package manager. In other cases, users will need to download and install beagle-lib manaully. Please be certain to their documentation and make sure that all listed prerequisites described therein are met prior to attempting compilation.
We have provided a Bash script which attempts to automate and verify the setup process. Please Note: as we are initialising environmental variables for your current shell, you must source this script (. bash_configure OR source bash_configure). Do not instead make it executable, as running it (./bash_configure) will not result in changes to your shell's environment.
. bash_configureUsers should pay attention to the output from the script. We attempt to highlight problems in red.
After initialisation, users may start sweeps using standard Nextflow invocation syntax or simply execute the chosen workflow scrip itself. Users will most likely wish to modify sweep to suit their own purposes, see below for an explanation of how a sweep is defined.
Any workflow can be started either using the following syntax:
nextflow run [options] <workflow>For instance, running Metagenomic-Hic stage 1 can be like so:
nextflow run chrcc-sweep.nfor even more simply, by treating any workflows as an executable:
./chrcc-sweep.nfIt is worth mentioning here that not all of our workflow scripts below are completely independent. For instance, Metagenomic Chromosome Conformation Capture is broken into three stages, with the second and third stages depending on the results of the previous stage. Therefore users must begin with stage 1 in this case.
Note: the nature of mixing concurrency and potentially resource hungry processes (such as genome assembly) can mean that a simple local execution strategy may result in resource starvation and subsequently premature program termination. It is recommended, in the long run, that users make use of a Nextflow supported distributed resource manager (DRM) such as SGE, SLURM, etc.
When a DRM is available as a target of execution, workflows can be easily submitted with only minimal configurational change to execute on potentially many physical nodes. Most frequently, this entails changes to the queue name for the target system. Even with only a single physical machine however, a well configured DRM can take better responsibility for managing available compute resources in completing a workflow's set of tasks.
Four options which users may frequently employ are (note: single hyphen) as follows.
Shorthand for including additional configuration details at runtime (perhaps when using different execution environments) and predefined in a nextflow configuration file under a given label [name]. For simplicity, these can be placed in the default nextflow.config and we have provided a few examples for a few different execution targets described below.
For many reasons, long running workflows may be interrupted either inadvertently or intentionally. For convenience and potentially a large time-saver, it is possible to restart or resume such jobs.
Limit concurrency to an integer number [Int] of simultaneous running tasks.
Specify a working directory other than the default work. This can he useful when separately running multiple workflows and you wish to inspect the contents of working directories without having to also determine which sub-folders pertain to which workflow.
Nextflow will describe its' base interface by invoking it alone on the command-line:
nextflowDetailed help with each command can be accessed as follows:
nextflow help [command]For even a relatively shallow parametric sweep, the sum total of computational resources required can be significant. Scheduling systems (SGE, PBS, SLURM, etc) are ideally suited to managing this task and Nextflow makes directing execution to them easy.
Specific scheduler submission details vary, as can the resources required for individual tasks. This information (executor, queue name, cpu, memory, disk) can be encapsulated either broadly in a config profile or per-process using process directives.
We have provided a few simple examples of such profiles within the default nextflow configuration file nextflow.config, which is automatically sourced by Nextflow at invocation.
We have attempted to use the simplest DRM-appropriate defaults, but queue names in particular frequently vary between deployments. Additionally, it is possible to specify many more criteria for your particular queuing system.
profiles {
standard {
process.executor = 'local'
}
sge {
process.executor = 'sge'
queue = 'all.q'
clusterOptions = '-S /bin/bash'
}
pbs {
process.executor = 'pbs'
queue = 'batch'
clusterOptions = '-S /bin/bash'
}
pbspro {
process.executor = 'pbspro'
queue = 'workq'
clusterOptions = '-S /bin/bash'
}
}
Submit to SGE queue manager
./chrcc-sweep.nf -profile sgeSubmit to a PBS queue manager
./chrcc-sweep.nf -profile pbsAll of our implemented workflows below accept the double-hyphen runtime option --debug.
Including this option at invocation time will test your environmental setup and the workflow processing logic but will not execute any actual bioinformatics tools. Workflows will execute very quickly in this mode and, if all is well, will complete without emitting an error. The mode creates only mock output files to meet inter-process dependencies. This mode is helpful when first configuring your system, particularly when configuring submission to a queue manager.
Nextflow writes to a hidden log file (.nextflow.log), which it manages in a rotation akin to Linux logrotate. If an error occurs during a workflow, it is very likely that Nextflow will expose it as a message to stdout, prompting the user to inspect the log file. In most cases, Nextflow will provide details on which task failed and its working directory. This directory should be the first port of call when troubleshooting a new problem. We would recommend inspecting both command.err (.command.err) and command.out (.command.out) to see if what went wrong can be ascertained. If not, inspect command.sh (.command.sh) and verify that the issued command is properly formed.
It is our experience that runtime errors are often caused by unset environmental variables. This can be as simple as inadvertantly switching shell terminals, where the second shell has not been configured. Therefore, even when you think you have done so already, please double check that you have initialized the environment as outlined above.
-
Unset environmental variables
-
Incorrect runtime architecture on execution host (requires x86_64)
-
Incorrect Java JVM. Please try using Java 8 or later.
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Out of storage space. Sweeps can occupy significant space.
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Exceeding CPU or other resource limits on execution host.
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Missing Python dependencies for invoked interpreter.
If using a Python interpreter other than the system default, make sure that the alternate's path (/home/foobar/bin) preceeds that of the system's default (/usr/bin).
e.g. PATH=/home/foobar/bin:/usr/bin
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No internet access.
If necessary, Meta-Sweeper uses Groovy Grape to automatically satisfy a few dependencies from internet repositories. Without access, compliation of our workflows will fail (see below).
In more severe cases, the Nextflow script might itself have failed to compile (groovy scripts are built at invocation time). In this situation, errors can be harder to decrypt and we would encourage users to contact us.
Primarily, a parametric sweep is defined by a set of variables which are sampled over a predefined range and granularity. In addition, there can be many other values held constant throughout the sweep, which are otherwise necessary and relevant to declare.
Effectively, the sweep is a sample-space where it is up to the user to decide on how finely the space is explored, whether the change between points is linear or otherwise. Once decided, the user declares the set of values for each variable in a simple configuration file.
Within the sweep configuration file, all varied parameters are grouped under the 'variables' label, while the remaining fixed values are collected under the 'options' label.
For the workflows implemented below, we have opted for a separate file for each worflow. Although not be strictly necessary, it permits keeping the size and complexity of the configuration file to a minimum, while more importantly avoiding unintended side-effects when changes with regard to one workflow are inadvertently taken up in another.
To tailor a sweep your preferences, users simply to extend or reduce the number of values taken on by any parameter. Note that at least one value must be defined for any variable used in a workflow. By defining a single value, a parameter is effectively fixed in the sweep. Ambition can quickly get the best of you and users should keep in mind that fine sampling of even a few parameters can lead to an exponential explosion in the full parameter space, potentially outstripping the computational resources at hand.
For any workflow, replicates are easily generated by defining multiple seeds.
How parameters vary in a sweep are defined in the configuration file. For the implemented workflows, there are a few more common parameters which we will now mention.
-
variables:This top most label is where swept parameters are defined. Please note: unfortunately, it is not possible to create new variables and have them dynamically added to a workflow. Only the values taken on in each case may be changed.-
seed:Where possible, all executables which rely on random seeds receive this value. Defining multiple seeds will result in replicates of the entire sweep. -
community:The critical entry defining the community to be analyzed. Here, a community is compose of an arbitrary number of phylogenetic clades. Each clade may use independent ancestral and donar (LGT) sequences. The per-clade phylogenetic trees are generated by birth-death simulation and pre-clade abundance profiles modeled as log-normal distributions. The size of each clade (number of taxa), the birth and death rates and log-normal parameters µ and σ are all user accessible. When multiple clades are defined, a final step merges them into a single normalized community. -
alpha:The branch length scale factor αBL controls the degree of evolutionary divergence between related taxa, with an recommended range of [0..1]. For a semi-qualitative perspective, evolutionary divergence measured in ANIb (average nucleotide identity by BLAST alignment) for an isotropic star topology will see ANIb < 85% (species divergence) when αBL ≅ 1 and ANIb > 95% (strain divergence) when αBL < 0.2 (Figure 1). -
xfold:Coverage, as measured in "times-genome-size" and used for workflows which make reference to conventional whole-genome shotgun sequencing of a community. For a community of 50 Mbp, xfold=10 would mean 500 Mbp of total sequencing or ~ 1.6 million 150 bp paired-end reads. -
n3c:For those workflows which deal with HiC sequencing, this variable defines the number of HiC read-pairs generated from a given community. In the case of 3C-contig graphs, 100,000 to 1 million pairs may prove sufficient for a simple community of four bacterial species. For larger or more complex communities, resulting in more fragmented WGS assemblies, extending as sweep beyond 1 million pairs is recommended.
-
Figure 1. Average nucleotide identity as a function of scale factor αBL
Taken from Metagenomic Chromosome Conformation Capture workflow, the following is an example of a sweep configuration file in YAML syntax.
variables:
# Level 0
# A complete sweep is run for each random seed defined here. In this case
# we will have 3 replicates of the entire sweep.
seed: [1, 2, 3]
# Level 1
# Communities are generated for each seed.
community: !com
name: trial
# A community of two clades.
# Clades can have any ancestral/donar sequence, but here they use the same.
# Altogether there will be 8 taxa in the community.
clades:
- !clade
prefix: clade1
ancestor: test/ancestor.fa
donor: test/donor.fa
ntaxa: 5
tree: {birth: 0.9, death: 0.5}
profile: {mu: 0.1, sigma: 1}
- !clade
prefix: clade2
ancestor: test/ancestor.fa
donor: test/donor.fa
ntaxa: 3
tree: {birth: 0.7, death: 0.3}
profile: {mu: 0.2, sigma: 1.5}
# Level 2
# Phylogenetic scale factor [0..1].
# The smaller this value, the more closely related
# taxa in each clade become.
alpha: [1, 0.5, 0.1]
# Level 3
# WGS sequencing depth, measured in times coverage
xfold: [1, 5, 10]
# Level 4
# The number of CCC read-pairs to generate
num_3c: [50000, 100000, 1000000]
options:
# How many samples. For most workflows, this is set to 1.
num_samples: 1
# Probabilties supplied to sgEvolver when generating community sequences.
evo:
indel_freq: 1e-4
small_ht_freq: 1e-4
large_ht_freq: 1e-4
inversion_freq: 1e-4
# WGS sequencing.
wgs:
read_len: 150
insert_mean: 450
insert_sd: 100
# 3C/HiC sequencing
ccc:
method: hic
enzyme: NlaIII
insert_mean: 500
insert_sd: 100
insert_max: 1000
read_len: 150
machine_profile: EmpMiSeq250
create_cids: true
# The output directory
output: outAt each point in the sweep, files which are considered outputs at various steps within a workflow are copied to the output folder [default: out]. For each such output file, the file name is used to pass-on information about the the parameters used at that given point in the sweep. This information is encoded as a structured string using two delimiters. These delimiters were chosen to avoid conflicts with system constraints, avoid unicode and be at least somewhat human readable.
This string is then prepended to the respective file name, which afterwards follows the regular convention of encoding information about the type of file.
Name/Value encoding:
Per-parameter syntax is delimited by a # character as follows:
[parameter_name]#[parameter_value]
Chaining parameters:
Multiple parameters are joined by the three character string -+-:
[parameter_name1]#[parameter_value1]-+-[parameter_name2]#[parameter_value2]
Example:
As an example, the following would be the file names for WGS reads R1/R2 pertaining to the sweep point seed=1, αBL=0.5, xfold=10:
seed#1-+-alpha#0.5-+-xfold#10.wgs.r1.fq.gz
seed#1-+-alpha#0.5-+-xfold#10.wgs.r2.fq.gz
In an effort to highlight the utility of Meta-Sweeper, we have implemented three workflows covering different metagenomic analysis topics we feel applicable to study by parametric sweep. In each case, the workflow definition can be adjusted through the respective configuration file, tailoring it to user requirements.
We encourage users to modify these examples for their own purposes.
This topic was our original motivation for creating Meta-Sweeper. The work culminated in the publication Deconvoluting simulated metagenomes: the performance of hard- and soft- clustering algorithms applied to metagenomic chromosome conformation capture (3C), where Meta-Sweeper represents a refinement of the methods employed in that work, allowing for its straightforward reproduction.
An added flexibility within this workflow is the ability to choose between the simulation of HiC or meta3C library preparation protocols. The most significant difference between the methods is that HiC employs a biotin affinity pull-down to greatly enrich for ligation products, while meta3C makes no attempt to do so. The outcome is that reads sequenced from a meta3C library will contain many conventional whole-genome shotgun (WGS) reads (>95%). Therefore, when simulating a meta3C workflow, we recommend users increase the requested sequencing yield (Eg. 10 * num_3c). At present, when running a meta3C simulation, the conventional purely WGS read-set is still created. In many real experiments, users might elect to produce only a single "meta3C enchanced" read-set, passing it to assembly as a plain WGS read-set.
The sweep is varied over five levels:
- Random seed
- Community structure
- Community divergence (αBL)
- WGS coverage (xfold)
- HiC ligation products (num_3c)
Sweep file: chrcc.yaml
The complete workflow has been broken into three stages, which we feel are often of separate interest.
-
Script: chrcc-sweep.nf
This first stage is responsible for the creation of communities, WGS and HiC read simulation, metagenome assembly and read mapping. The results from this stage are copied to the output folder [default: out]. How each each parameter should vary within the sweep can be adjusted in the configuration file. We would recommend users consider storage and CPU requirements prior to expanding the size of the sweep.
Note: the definition as it stands in only toy-like in size.
-
Script: chrcc-cluster.nf
After stage 1 has completed, stage 2 out clustering of the 3C-contig graphs resulting from stage 1. At present, clustering is performed with the algorithms: Louvain-hard, Louvain-soft and OClustR. Afterwards, performance and quality metrics are applied. The BCubed external metric is used to assess the performance of each algorithm relative to the ground truth, while simple statistics for assembly (N50, L50) and graphs (size, order) are compiled alongside an entropic measure of graphical complexity (HL)
-
Script: chrcc-aggregate.nf
Acting effectively as a reduce function, this is the simplest stage. Here the results from potentially many permuted outcomes are collected and aggregated into a single results file all_stats.yaml.
This results file (e.g.
out/all_stats.yaml) is organized as a list, where each element encompasses both the input parameters and resulting outcomes from one sweep point. The elements are stored as associative collections, one per sweep point, where validation results from stage 2 are grouped by the statistical test performed.The file can be deserialized to an object within any language where support for YAML exists (i.e. Python, Java & Groovy, Ruby, C++, etc.) If users do not wish to deal with a serialized object, the result file can be converted to a flat table using the script
tabular_results.py. When converted to tabular form, rows represent the results for each point in the sweep.
The serialized YAML object contains the following entries:
- params - the set of input sweep parameters
- asmstat - common assembly statistics
- bc - the weighted BCubed external index (see manuscript)
- geigh - a entropic measure of graphical complexity (see manuscript)
- gstat - common graph statistics
Example: two sweep points from all_stats.yaml
- params: {seed: '2', alpha: '1', xfold: '1', n3c: '5000', algo: louvsoft}
asmstat: {L50: 12, N50: 455}
bc: {completeness: 0.964, f: 0.141, pre: 1.0, rec: 0.0757}
geigh: {method: eigh, value: null}
gstat: {density: 0.0741, inclusives: 28, isolates: 0, mean_deg: 2.0, median_deg: 2, modularity: 0.964, order: 28, size: 28}
- params: {seed: '3', alpha: '1', xfold: '2', n3c: '5000', algo: louvsoft}
asmstat: {L50: 130, N50: 568}
bc: {completeness: 0.953, f: 0.0152, pre: 1.0, rec: 0.008}
geigh: {method: eigh, value: -0.0}
gstat: {density: 0.006, inclusives: 342, isolates: 0, mean_deg: 2.006, median_deg: 2, modularity: 0.997, order: 342, size: 343}which when converted to tabular format would look like:
,params-xfold,params-alpha,params-n3c,params-seed,params-algo,asmstat-L50,asmstat-N50,gstat-mean_deg,gstat-density,gstat-modularity,gstat-median_deg,gstat-inclusives,gstat-isolates,gstat-order,gstat-size,geigh-method,geigh-value,bc-pre,bc-rec,bc-completeness,bc-f
0,1,1,5000,2,louvsoft,12,455,2.0,0.0741,0.964,2,28,0,28,28,eigh,,1.0,0.0757,0.964,0.141
1,2,1,5000,3,louvsoft,130,568,2.006,0.006,0.997,2,342,0,342,343,eigh,-0.0,1.0,0.008,0.953,0.0152
A test case for metagenome deconvolution by non-negative matrix factorisation of a time-series data-set.
This workflow is implemented as a single script. Internally, the stages of execution involve:
- Community creation
- WGS read simulation
- WGS read mapping
- NNMF based deconvolution
The sweep is varied over four levels:
- Random seed
- Community structure
- Community divergence (αBL)
- WGS coverage (xfold)
This workflow does not involve HiC sequencing data.
Script: timeseries-deconvolute.nf
Sweep file: timeseries.yaml
The final outcome of the deconvolution process for each sweep point can be found in the files following the naming strategy: ${key}.truth.report.txt. Here, the variable ${key} represents the structured string encoding information about the parameters used at a given point in the sweep. This is explained above
E.g. For seed=1, αBL=0.5 and xfold=10, the report file would be named:
seed#1-+-alpha#0.5-+-xfold#10.truth.report.txt
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metaART – Simulation of whole-genome shotgun paired-end reads from communities.
In essence, MetaART is a simple wrapper of the art_illumina binary from the ART suite (Huang et al, 2011). Art_illumina supports the simulation of Illumina-based paired-end WGS sequencing and supports many machine types and sequencing modes. Our extension allows the association of relative abundances with the set of supplied reference sequences as as to better model the outcome of metagenomic sequencing or multi-chromosomal genomes with varying copy-number.
Profiles can be supplied either in the form of an explicit table or drawn at random from one of three distribution at runtime (equal, uniform random, log-normal). Reads from each reference are then generated to the required depth (a product of the requested coverage and the genome's abundance). The combined result is returned as a single read-set. Simulations are initialised via a global random seed, which is also passed on to the art_illumina binary. Although not all of art_illumina's options have been passed on to the user: read length, insert length and standard deviation can be user set, as well as machine type.
Example Uses
For a predefined abundance profile
abdn.tsv, 10x overall coverage of the reference multifastarefseq.fastaand with the result written tooutdir:metaART.py -S 1234 --profile abdn.tsv --read-len 150 --insert-len 500 --insert-sd 100 --max-coverage 10 refseq.fasta outdir
Or the same job but with runtime generated abundances using a log-normal distribution:
metaART.py -S 1234 --dist lognormal --lognorm-mu 0.1 --lognorm-sigma 1.0 --read-len 150 --insert-len 500 --insert-sd 100 --max-coverage 10 refseq.fasta outdir
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sim3C – Simulation of HiC/3C read-pairs.
Analogous to the well established process of simulating whole-genome shotgun reads,
sim3C.pysimulates read-pairs as if generated from a sequencing library prepared using a HiC/3C methodology.Experimental noise is considered both in terms of Illumina-based sequencing error and that inherent in the production of HiC/3C library generation. Sequencing error is afforded by a reimplementation of
art_illumina(Huang et al, 2011) in Python (Art.py). HiC/3C noise, in the form of spurious ligation products, is modeled as the uniformly random association of any two sites across the entire extent of the source genome(s), the rate of which is user controlled.To support community sampling (metagenomes), an abundance profile is supplied along with the set of reference sequences at runtime. The profile can either take the form of an explicit table or be drawn at random from a user chosen probability distribution (equal, uniform random or log-normal).
The tool conceptualises the process of HiC/3C read-pair generation, as a series of conditional probabilities. For intra-chromosomal read-pairs, genomic separation is constrained to follow an empirically determined long-tailed composition of the geometric and uniform distributions. For inter-chromosomal (but same cell) pairs, genomic position is constrained only by proximity to a restriction site. Lastly, inter-cellular read-pairs are not considered.
Fine scale structurally related features that have been observed via contact maps in real experiments (Tung et al, 2013) are also reproduced in our simulation. Namely, contacts between the two arms of the chromosome and chromosomal interacting domains (CID). Within bacterial HiC contact maps, inter-arm contacts are responsible for the fainter anti-diagonal observed (y=-x rather than y=x), while the tightly folded CID domains act to modulate contact frequencies over their local extent, resulting in blocks of density. In our simulation, CIDs are randomly generated at runtime using the supplied seed, and the strength of their effect adjusted or disabled by the user.
Pseudocode
If a spurious event: unconstrained pairing of any two sites across entire (meta)genome Else if an inter-chromosomal event: unconstrained positions on Chr_A, Chr_B from Genome_N Else is an intra-chromosomal event: contrained positions x1, x2 ~ g(x) on Chr_AWhere g(x) reflects both the empirical constraint on distance and fine-scale features mentioned above.
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louvain_cluster – Louvain community detection based graph clustering. Both traditional hard-clustering solutions of the best partition and a naive soft-clustering option (DeMaere and Darling, 2016).
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oclustr – An implementation of the OClustR graph hard-clustering algorithm (Pérez-Suárez et al, 2013).