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REAT - Robust and Extendable eukaryotic Annotation Toolkit

REAT is a robust easy-to-use genome annotation toolkit for turning high-quality genome assemblies into usable and informative resources. REAT makes use of state-of-the-art annotation tools and is robust to varying quality and sources of molecular evidence.

REAT provides an integrated environment that comprises both

  • a set of workflows geared towards integrating multiple sources of evidence into a genome annotation, and
  • an execution environment (Docker, Singularity) for these workflows.

Getting Started

To configure a development environment for REAT you need:

  • the REAT source code
  • Software dependencies
  • A WDL compatible workflow engine (REAT development is mainly geared towards using Cromwell)

To obtain the REAT source code, please:

git clone https://github.com/ei-corebioinformatics/reat

While an exhaustive list of software dependencies is available in the appendix, a simpler way of obtaining all the required binaries for REAT is to build a Singularity container based on the definition file (reat_singularity.def) included in the repository.

singularity build reat.img reat_singularity.def

Cromwell can be obtained from the original repository at https://github.com/broadinstitute/cromwell/releases.

Prerequisites

For ease of development, Singularity is recommended. In case this is not available (you are working on MacOS and haven't got https://sylabs.io/guides/3.5/admin-guide/installation.html#mac available), please follow the instructions in the REAT/reat_singularity.def for building and installing the binary dependencies.

Java 8

Singularity (optional)

Cromwell >= v48

You will need to download this from the Cromwell repository

Installing

First obtain the source code using

git clone https://github.com/ei-corebioinformatics/reat
cd reat

You should now have the latest development version, in which you will find the reat_singularity.def. This allows you to build a Singularity container.

We also provide a conda environment which you can use to install dependencies if you prefer.

conda env create -f reat.yaml --name reat

Alternatively, install the following software dependencies on your system, so that the executables are available in your $PATH environment variable.

  • DIAMOND - 0.9.31
  • BioPerl
  • FullLengtherNext - 1.0.1
  • Install BLAST - 2.7.1
  • Install MagicBlast - 1.5.0
  • LibDeflate - master
  • HTSLib - 1.9
  • Samtools - 1.9
  • BCFTools - 1.9
  • BAMtools - 2.5.1
  • gclib - 54917d0
  • gffread - ba7535f
  • GMAP - 2019-02-15
  • MiniMap2 - 2.17
  • GenomeTools - 1.5.10
  • HISAT2 - 2.1.0
  • STAR - 2.7.3a
  • seqtk - master
  • Stringtie2 - v2.1.1
  • Scallop - 0.10.4
  • Scallop-lr - 0.9.2
  • Prodigal - 2.6.3
  • Transdecoder - 5.5.0
  • Portcullis - 1.2.2
  • Mikado - 2.0

Instructions for building each of the tools can be found in the reat_singularity.def.

Please install reat to ensure the scripts are findable during execution. To install, simply use from your current pip/conda environment:

pip install ./reat

Running REAT

Cromwell supports a range of execution backends such as cloud services and batch schedulers. These can be configured in files such as those present in the cromwell_configuration directory. For more examples you can also visit https://github.com/broadinstitute/cromwell/tree/develop/cromwell.example.backends

Local

REAT finds and executes a binary named cromwell which can be configured by placing a file similar to:

#!/bin/sh
java -Dconfig.file=reat/cromwell_configuration/local.conf -jar cromwell.jar

Where local.conf is the file in this repository under cromwell_configuration and cromwell.jar is a cromwell release jar file (https://github.com/broadinstitute/cromwell/releases)

Finally, for the homology module you can run:

reat --workflow_options_file workflow_options/options.json --computational_resources compute_inputs.json homology <homology_args>

And, for the transcriptome module:

reat --workflow_options_file workflow_options/options.json --computational_resources compute_inputs.json transcriptome <transcriptome_args>

SLURM

To run REAT in a SLURM backend the cromwell instance needs to be configured using the configuration file equivalent for your environment to cromwell_configuration/slurm.conf. This file contains the configuration required by Cromwell to execute jobs using a SLURM backend. It includes details such as how to submit, check and kill a job running on a SLURM backend.

Running reat under slurm requires changing the -Dconfig.file argument from the local example of our cromwell command to:

#!/bin/sh
java -Dconfig.file=reat/cromwell_configuration/slurm.conf -jar cromwell.jar

Workflow

Transcriptome Workflow

Alt text

Homology Workflow

Alt text

Prediction Workflow

Alt text

Releasing new versions

To create a new release, make sure your repository is in a clean state and please use:

bumpversion minor --verbose --tag --message "Minor version bump" --tag-message "Minor version bump

List of changes (note that this message will be used in the Release)
Finally you may want to update the release message by editing it directly

The above will bump a minor version (options are major, minor and patch), create a tag with the message above, trigger the github actions for checking the commit and generating a new github release.