Description:

sequanademultiplex 1.5.2

This is is the demultiplex pipeline from the Sequana projet

Overview:
Runs bcl2fastq on raw BCL data and creates plots to ease the QC validation

Input:
A valid Illumina base calling directory and sample sheet file

Output:
An HTML report, a set of PNG files and the expected FastQ files

Status:
production

Wiki:
https://github.com/sequana/demultiplex/wiki

Documentation:
This README file, the Wiki from the github repository (link above) and https://sequana.readthedocs.io

Citation:
Cokelaer et al, (2017), ‘Sequana’: a Set of Snakemake NGS pipelines, Journal of Open Source Software, 2(16), 352, JOSS DOI https://doi:10.21105/joss.00352



Installation
Intall the sequana_demultiplex package as follows:
pip install sequana_demultiplex


Usage
sequana_demultiplex --help
sequana_demultiplex --working-directory DATAPATH --bcl-directory bcldata --sample-sheet SampleSheet.csv --merging-strategy merge
The –bcl-directory option indicates where to find your raw data, the sample-sheet
expects the SampleSheet to be compatible with IEM software. The –merging-strategy can
be set to none or merge. The merge option merges the lanes, which is
useful for e.g. NextSeq sequencers.
This creates a directory fastq. You just need to execute the pipeline:
cd demultiplex
sh demultiplex.sh # for a local run
These commands launch a snakemake pipeline. If you are familiar with snakemake, you can retrieve the demultiplex.rules and config.yaml files and then execute the pipeline yourself with specific parameters:
snakemake -s demultiplex.rules --cores 4 --stats stats.txt \
--wrapper-prefix https://raw.githubusercontent.com/sequana/sequana-wrappers/"
You may also use sequanix for a graphical interface.
Would you need to merge the lane, please add the –merging-strategy argument
followed by merge:
sequana_demultiplex --bcl-directory bcl_data --merging-strategy merge --sample-sheet SampleSheet.csv


Requirements
This pipeline requires the following third-party tool(s):

bcl2fastq 2.20.0

This software has an end-user license agreement (EULA). Given the EULA details
of this software, it cannot be distributed according to ` Illumina license <https://support.illumina.com/content/dam/illumina-support/documents/downloads/software/bcl2fastq/bcl2fastq2-v2-20-eula.pdf>`_
Therefore, you should install it yourself. On cluster facility, you may ask to
your system administator. For instance:
module load bcl2fastq/2.20.0
For the same reason you cannot find it on community such as bioconda or docker (aug 2020).
So, you will need to download the code yourself. The easiest is to download the
RPM from Illumina
and accept the agreements. Install it using the RPM if you have a debian-like system:
rpm install file.rpm
If you do not have a debian system, you can look at https://damona.readthedocs.io where we provide
a singularity recipes to build an image from your own rpm. Recipes can be found
here.


Details

This pipeline runs bcl2fastq 2.20 and creates a set of diagnostics plots to help
deciphering common issues such as missing index and sample sheet errors.


Rules and configuration details
Here is the latest documented configuration file
to be used with the pipeline. Each rule used in the pipeline may have a section in the configuration file.


Changelog


Version
Description



1.5.2

rename requirements.txt into tools.txt; update __init__



1.5.1

switch working directory to fastq instead of demultiplex(regression)



1.5.0

Uses click and new sequana_pipetools, add multiqc



1.4.0

Implement demultiplexing of single cell ATAC seq data with
cellranger.



1.3.1

use sequana_wrappers version in the config file



1.3.0

use latest sequana-wrappers to benefit and graphivz apptainer



1.2.1

Update CI action and use new sequana_pipetools v0.9.0



1.2.0

stable release with cleanup of the setup and README



1.1.3

add the –mars-seq option that fills the config automatically



1.1.2

fix the none_and_force merging strategy option



1.1.1

fix a regression bug



1.1.0

Uses new sequana-wrappers repository



1.0.5

Fix regression bug to cope with new snakemake API
Compatibility with sequanix GUI



1.0.4

Better HTML report with updated images.
validate the SampleSheet when using sequana_demultiplex and/or the
pipeline
Add error handler from sequana_pipetools
save all undetermined barcodes (not just first 20)
No changes to the UI
technically, the input_directory option is now in a section so that
it can be used in Sequanix



1.0.3

remove check_samplesheet and fix_samplesheet modules now in sequana
check sample sheet but do not fail. Instead, informing users that
there is an error and suggest to use ‘sequana samplesheet
–quick-fix’



1.0.2
Use ‘sequana samplesheet –check ‘ command instead of deprecated
sequana_check_sample_sheet command

1.0.1
change some default behaviour:

write_fastq_reverse_complement is now set to False by default
like bcl2fastq
The –no-bgzf-compression option is changed into
–bgzf-compression. We do not want this option by default.
The –ignore-missing-bcls option is changed into
–no-ignore-missing-bcls so as to ignore missing bcls by default
keep this option as a flag and keep same behaviour
Fix HTML syntax



1.0.0

stable version pinned on sequana libraries



0.9.11

fix label in plot_summary,
add new plot to show reads per sample + undetermined
add two tools one to check the samplesheet called
sequana_sample_sheet and one called sequana_fix_samplesheet. The
former is now inside the pipeline as well and when creating the
pipeline
set –write_reverse_complement to False by default
remove the –ignore-missing-control which is deprecated anyway



0.9.10

implement the new option –from-project, add missing MANIFEST



0.9.9

simplification of the pipeline to use sequana 0.8.4 to speed up
the –help calls.
include a summary HTML report



0.9.8

fix typos



0.9.7

Use new release of sequana_pipetools
set matplotlib backend to agg
include a simple HTML report



0.9.6

Handle different RunParameter.xml name (NextSeq vs HiSeq)



0.9.5

Fix a regression bug due to new sequana release. We do not check
the input file (fastq) since this is not a sequence analysis
pipeline
Check whether it is a NextSeq run. If so, merging-strategy must be
set to ‘merge’. Can be bypassed using –force



0.9.4

Check the presence of the bcl input directory and samplesheet.
More help in the –help message.
add –sample-sheet option to replace –samplesheet option
Fix the schema file
Check for presence of RunParameters.xml and provide information
if merging-stratgy is set to None whereas it is a NextSeq run



0.9.3
Fix regression bug

0.9.2
remove warning due to relative paths.

0.9.1
Make the merging options compulsory. Users must tell whether they
want to merge the lanes or not. This avoid to do the merging or not
whereas the inverse was expected.

0.8.6
Uses 64G/biomics queue and 16 cores on a SLURM scheduler





Contribute & Code of Conduct
To contribute to this project, please take a look at the
Contributing Guidelines first. Please note that this project is released with a
Code of Conduct. By contributing to this project, you agree to abide by its terms.

License

For personal and professional use. You cannot resell or redistribute these repositories in their original state.

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