Dockerized Genomic Processing Pipelines

Genomic data processing pipelines written in Workflow Definition Language (WDL), making use of Docker containers to ensure reproducability. The Docker containers used have been curated for usage in the Faryabi Lab processing pipelines, and can be found on Dockerhub.

Setup

Clone this repository to your local machine

git clone https://github.com/faryabiLab/dockerize-workflows

Dependencies

Ensure that the latest cromwell engine (.jar file) and womtools (.jar) is installed from their GitHub repository.
Have Docker installed on your machine.

Sample Sheet

Each run of a workflow must be accompanied by a samplesheet.tsv - a single-column tab-separated file that specifies the prefixes ('sample names') of each fastq file to be processed.
A note on fastq naming - Ensure that your paired-end fastq files follow the naming convention XYZ_R1.fastq.gz / XYZ_R2.fastq.gz, where XYZ represents this file's sample name. Single-end fastqs must be named as XYZ.fastq.gz. To make a sample sheet, simply run make_samplesheet.sh found in utils/:

• Paired-end: ./make_samplesheet.sh -p -d /path/to/fastq_dir
• Single-end: ./make_samplesheet.sh -d /path/to/fastq_dir

This will create a samplesheet.tsv in your current working directory.

Options file

Cromwell allows the user to pass in an options file via the -o/--options argument, which grants control over the pipeline's output locations. This file's format is that of standard JSON, and the keys made readily available in this repository are:

• final_workflow_outputs_dir - Directory to which the final workflow outputs will be copied
• final_call_logs_dir - Directory to which the final workflow logs will be copied There are more options available, you can find them in the Cromwell documentation.

Docker Containers

By default, the pipelines are configured to run each step within a specified docker container, all of which are hosted on the Faryabi Lab Dockerhub. Workflow input files contain a Dockerhub_Pull variable for each step that can be used to point to another Docker container, although this could be risky.

Cromwell Configuration

Within the cromwell_configs directory is a Cromwell config file which instructs the engine to use Docker as backend. The only option that can be tweaked in this file is the concurrent_job_limit variable: this controls the number of jobs that can be running at once (default = 10).

Call-Caching

A desirable feature for many workflows is the ability to identify steps, or "jobs", that have already been run and essentially "skip" that step, saving both time and computing resources. In this reporisoty, Cromwell is configured to use a local MySQL database (which runs from a Docker container) called PipelineDatabase, to store intermediate files to determine the steps that have been run. If you want to use a different SQL, or any, database, the database block must be edited accordingly in the Cromwell configuration file, else, the config will attempt to log in to the MySQL server specified. By default, the database block is configured to work with a MySQL Docker container that can be initialized via the following command:

docker run \
-p 52000:3306 \
--name PipelineDatabase \
-e MYSQL_ROOT_PASSWORD=@noah1234 \
-e MYSQL_DATABASE=PipelineDatabase -e MYSQL_USER=pipeline \
-e MYSQL_PASSWORD=Run@pipelines9061 \
-d mysql/mysql-server:latest

Usage

Inputs

The /workflows directory houses all by-assay pipelines. In each subdirectory, there are several files:

• *.wdl - The main workflow file that will be run.
• imports.zip - A zipped directory of all imports needed for this pipeline. This is important for running your pipeline successfully.
• *_inputs.json - A JSON file with all relative input options.
• options.json - A JSON file with paths to output locations for workflow results and logs.

In the inputs JSON, you will find command-specific parameters (i.e. for trimming, alignment, etc.) as well as common inputs needed for every pipeline:

• paired - A boolean value indicating whether the experiment was paired-end (true) or single-end (false).
• project_out_dir - Directory where output will be (a directory is created here for every sample name).
• fastq_dir - Path to dorectory with all .fastq.gz files.
• sampleList - Path to your previously created samplesheet.tsv
• star_index/BWA_index - Path to alignment index files.
• chromNoScaffold - A Path to a 3-column BED file with the chromosome/contig regions you would like to keep in your output.
• GeneAnnotationFile - Path to .gtf file.
• chromosome_sizes - A standard BEDtools chromosome size file.
• blacklist - A 3-column BED file containing areas to be removed from output.

Running a Workflow

Once the configuration is complete, there are 2 options available to you to run your pipeline:

1. Run in local mode

• Cromwell can run "on-the-fly", without the need to configure a server. To do this, use the run subcommand as such:

java \
  -Dconfig.file=/path/to/cromwell_config \
  -jar /path/to/cromwell.jar run \
  -i workflow_input.json \
  -o options.json \
  -p imports.zip \
  workflow.wdl

• This will startg a Cromwell instance, run your workflow, and then exit upon completion.

2. Submit to a Cromwell server

• First, if a Cromwell server isn't already running, start one:

java -Dconfig.file=/path/to/cromwell_config -jar /path/to/cromwell.jar server

• Then, submit your workflow to the server via the submit command along with the host address, imports.zip, inputs.json, and your workflow's WDL file:

java \
  -Dconfig.file=/path/to/cromwell_config \
  -jar /path/to/cromwell.jar submit \
  -h http://<host ip>:<port> \
  -p imports.zip \
  -o options.json \
  -i inputs.json \
   workflow.wdl

• Navigate to 0.0.0.0:8000 to access the GUI, where different aspects of the workflow can be monitored via the REST API.
  • A useful tool is the timing diagram tool which displays a graphical representation of the workflow's progress, broken down by sample and job. It can be accessed with the REST API via 0.0.0.0:5200/api/workflows/v1/<workflow ID>/timing \

Of course, you can find out more about Cromwell's capabilities in the Official Cromwell Documentation.