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pgx_dnaseq - Automatic NGS pipeline

Version 0.9

pgx_dnaseq is an automatic pipeline to perform high-throughput sequencing analysis. It also provides useful tools such as coverage_graph.py and read_quality_graph.py.

Dependencies

The tool requires a standard Python installation (>=3.4) with the following modules:

  1. numpy version 1.8.1 or latest
  2. pandas version 0.13.1 or latest
  3. ruffus (version 2.6b or latest and available here)
  4. matplotlib version 1.3.1 or latest

The tool has been tested on Linux only, but should work on MacOS operating systems as well.

Installation

First, create a directory where the Python virtual envirnoment will be installed.

$ mkdir -p ~/softwares/Python-3_virtualenv
$ pyvenv ~/softwares/Python-3_virtualenv

Then, activate the newly created virtual environment:

$ source ~/softwares/Python-3_virtualenv/bin/activate

Before installing the pipeline, install numpy:

$ pip install -U numpy

Finally, install the pipeline (it will install all the remaining requirements):

$ pip install -U pgx_dnaseq-0.9.tar.gz

Usage

Pipeline

This is the usage of the automatic pipeline.

$ execute_pipeline.py --help
usage: execute_pipeline.py [-h] [-v] [-i FILE] [-p FILE] [-t FILE] [-d]
                           [-n INT] [--preamble FILE] [-f FORMAT]

Execute a NGS pipeline (part of pgx_dnaseq version 0.9).

optional arguments:
  -h, --help            show this help message and exit
  -v, --version         show program's version number and exit

Pipeline Configuration:
  -i FILE, --input FILE
                        A file containing the pipeline input files (one sample
                        per line, one or more file per sample.
                        [input_files.txt]
  -p FILE, --pipeline-config FILE
                        The pipeline configuration file. [pipeline.conf]
  -t FILE, --tool-config FILE
                        The tools configuration file. [tools.conf]
  -d, --use-drmaa       Use DRMAA to launch the tasks instead of running them
                        locally. [False]
  -n INT, --nb-process INT
                        The number of processes for job execution (allow
                        enough if '--use-drmaa' option is used since you want
                        at least one job per sample). [1]
  --preamble FILE       This option should be used when using DRMAA on a HPC
                        to load required module and set environment variables.
                        The content of the file will be added between the
                        'shebang' line and the tool command.

Pipeline Flowchart:
  -f FORMAT, --format FORMAT
                        The format of the pipeline flowchart [pdf]

When using DRMAA on a HPC requiring module to be loaded (e.g. java), don't forget to set the preamble option. Be careful though on what is written in the preamble file, since it is written as is in the temporary script used by DRMAA. Also, if using a Python virtual environment, don't forget to activate it in the preamble file in order to use provided Python script.

Automatic reporting

The package also provide automatic reporting, using the following script.

$ automatic_report.py --help
usage: automatic_report.py [-h] [-v] [--debug] [-i FILE] [-p FILE] [-r STRING]
                           [-o FILE]

Generate automatic report (part of pgx_dnaseq version 0.9).

optional arguments:
  -h, --help            show this help message and exit
  -v, --version         show program's version number and exit
  --debug               Set the log level to debug.

General Options:
  -i FILE, --input FILE
                        A file containing the pipeline input files (one sample
                        per line, one or more file per sample
                        [input_files.txt]
  -p FILE, --pipeline-config FILE
                        The pipeline configuration file [pipeline.conf]

Report Options:
  -r STRING, --run-name STRING
                        The Sequencing run name [Sequencing_Run]
  -o FILE, --output FILE
                        The name of the final report [dna_seq_report.pdf]

Provided quality script

The read_quality_graph.py script produces a pretty read base quality distribution plot. Here is its usage:

$ read_quality_graph.py --help
usage: read_quality_graph.py [-h] [-v] [--debug] [--log LOGFILE]
                             [-i FILE FILE] [-o FILE] [-t TITLE]

Produces a pretty read base quality distribution plot (part of pgx_dnaseq
version 0.9).

optional arguments:
  -h, --help            show this help message and exit
  -v, --version         show program's version number and exit
  --debug               Set the logging to debug
  --log LOGFILE         The log file [read_quality_graph.log]

Input Files:
  -i FILE FILE, --input FILE FILE
                        The input FASTQ or FASTQ.GZ files

Result File:
  -o FILE, --output FILE
                        The name of the output file [read_quality.pdf]
  -t TITLE, --title-prefix TITLE
                        The title of the plot []

The coverage_graph.py script plots NGS coverage using samtools mpileup. Here is its usage:

$ coverage_graph.py --help
usage: coverage_graph.py [-h] [--version] [--samtools-exec PATH]
                         [--depth-file FILE [FILE ...]] [--bam BAM [BAM ...]]
                         --bed BED [-q INT] [-Q INT] [-d INT]
                         [--max-depth INT] [-o FILE]

Plots NGS coverage (part of pgx_dnaseq version 0.9).

optional arguments:
  -h, --help            show this help message and exit
  --version             show program's version number and exit
  --samtools-exec PATH  The PATH to the samtools executable if not in the
                        $PATH variable

Input Files:
  --depth-file FILE [FILE ...]
                        Results from this script (to redo the plot faster)
                        (one or more, separate by spaces)
  --bam BAM [BAM ...]   Input BAM file(s) (one or more, separated by spaces)
  --bed BED             BED file to restrict to targeted regions

MPILEUP Options:
  -q INT                skip alignments with mapQ smaller than INT [0]
  -Q INT                skip bases with baseQ/BAQ smaller than INT [13]
  -d INT                max per-BAM depth to avoid excessive memory usage
                        [250]

Plotting Options:
  --max-depth INT       The maximal depth to plot (in order to zoom in the
                        plots) [None]

Output Options:
  -o FILE, --out FILE   The name of the output file [depth]