Toolbox for generic NGS analyses

MANUAL for TOGGLE v0.3 for ON-THE-FLY pipeline creation


The TOGGLe v0.3.x version allows users to create their own customized pipelines. You can modify not only the different options for each software but also define dedicated pipelines for your analyses.

You can therefore switch, add and remove steps, start from FASTA/FASTA.GZ, FASTQ/FASTQ.GZ, SAM/BAM, BED, GFF or VCF/VCF.GZ files, ask for individual treatments only, individual then common (such as individual mapping followed by common calling), or even only common treatments.

Input files naming convention

TOGGLE will automatically assign sample name (readgroup) based on the input file name structure, picking up the “text” before the first dot.

  • individual1_1.fastq will be understood as individual1
  • mapping1.sam will be understood as mapping1
  • myVcf.complete.vcf will be understood as myVcf

For Fastq files, only one underscore (‘_’) is allowed, before the direction of sequences ( _1, _2 or _R1, _R2), just before the extension (.fastq).

Please use only UTF-8 standard symbols, no weird characters or space, pipe, tilde or any type of commas.

Launching an analysis

The current version is based on the script -d|--directory DIR -c|--config FILE -o|--outputdir DIR [-r|--reference FILE] [-k|--keyfile FILE] [-g|--gff FILE] [-nocheck|--nocheckFastq] [--help|-h]
Required named arguments:  
-d / –directory DIR: a folder with raw data to be treated (FASTA/FASTA.GZ, FASTQ/FASTQ.GZ, SAM, BAM, BED, GFF, VCF/VCF.GZ)
-c / –config FILE: generally it is the software.config.txt file but it can be any text file (Unix format) structured as shown below .
-o / –outputdir DIR: the current version of TOGGLe will not modify the initial data folder but will create an output directory with all analyses in. This module must be empty (TOGGLe will stop if not).
Optional named arguments:  
-r / –reference FILE: a reference FASTA file to be used. (1)
-g / -gff FILE: a GFF file to be used for some tools . Be careful the gff name must be different than the FASTA.
-k / –keyfile FILE: a keyfile use for demultiplexing step.
-nocheck / –nocheckFastq: by default TOGGLe checks if given formats for input files are correct. This option allows to skip this step.
-report / –report: generate pdf report (more info)
-h / –help: show help message and exit

(1): If no reference indexes exist, they will be created accordingly to the pipeline requested indexes. If they exist, they will not be re-created UNLESS the pipeline order (see below) expressively requests it (to update the index e.g.)

All paths (files and folders) can be provided as absolute (/home/mylogin/data/myRef.fasta) or relative (../data/myRef.fasta).

What will you have in results ?

TOGGLe will generate an output folder containing different files and subfolders, as follows:


The final results are contained in the finalResults folder. TOGGLe will also copy the software config file corresponding to the analysis, in order users can recover their options. The output folder contains all sub analyses, i.e. the individual analyses or intermediate data.

Writing a whole configuration file to create a pipeline

The script will use the configuration file informations (here named as software.config.txt file) to create specific pipeline scripts in the output folder, called for individual treatments (individual mapping e.g.) and for global treatment (global SNP calling e.g.).

The order will be checked before creating the scripts, to control that the output files from the step n-1 will be accepted as input for the step n.


Providing an order

The order of a specific pipeline is provided in the software.config.txt as the software order

Thus, if you provide options such as:

1=bwa aln
3=samtools view

You will obtain a pipeline taking FASTQ files as input (plain text of gzipped), aligning them upon a given reference using bwa aln/sampe, and finally cleaning the resulting sam and providing a BAM file (if samtools view option are as such).

Note that the way you write the name of the step is not really important, e.g. it is not case-sensitive, and can have space between the different words. A dedicated module will adjust everything.

You can start from any type of format recognized by TOGGLE, and from any step. Thus, if you have a raw VCF file you can start from this one at step one and clean it as follows (even if your VCF contains informations for multiple individuals):


BE CAREFUL: You can comment any step using a ‘#’ symbol before it (ex #3=bwaAln) to avoid to retype all numbers. However, such a comment will provoke errors in cleaning and compressing steps. The best approach is to provide a list of following numbers (e.g. 1, 2, 3, 4 and not 1, 3, 4). This is also true for steps higher than 1000.

Same software repeated multiple times

If you want to adress multiple times the same step BUT with different configurations (e.g. a first samtools view to obtain a BAM then a second samtools view to clean this BAM using the -f option), you have to significate it in the order key as follows:

1=bwa aln
3=samtools view 1
4=samtools view 2

In the same time you have to provide the same informations in your configuration:

$samtoolsView 1
$samtools View 2
-f 0x02

BE CAREFUL: in such multiple times repeated software, you have to put a space between the software name and the step (ex samtoolsview 1 and samtoolsView 2) in the order as well as in the options !!

Giving a common step to all individuals (multiple entry files)

If you want for instance to map all individuals (multiple files) separately then perform a common SNP calling, please add a step number higher than 999:

3=picardTools SortSam

This pipeline will map FASTQ then sort per coordinates the SAM for each individuals, and then will launch a common step for all as a global SNP calling. You can add after this calling other steps (1001=gatkVariantFiltrator for example).

Warnings: if you want to map and call SNP from only one sample, it is not necessary to call a 1000+ step (here gatkUnifiedGenotyper would be then the 4th step).

Starting only as a common treatment

If you want only a global treatment for multiple files (you may have all your formatted BAM and you want to perform the calling and subsequent steps), you can create the following pipeline:

1000=gatk UnifiedGenotyper
1001=gatk VariantFiltrator

The pipeline will treat the data as a global pipeline in this case, without separating the individual files.

Sending software parameters

Any software configuration will start as follows:

 $First Software

 $Second Software

You can address any option to any given software (as soon as the given option exists for the given software ^^ ) as follows:


You can also write as:


The software name is not case sensitive and the subprogram can be “glued” to the name (bwaALN is recognized, as well as bwa aln).

If your option has an equal symbol within, such as -l hostname=MyNode, you have to write the option as follows:

-l hostname=MyNode

Cleaning steps

BETA FEATURE: Using the cleaning steps may impair the execution of some steps. Use it carefully.

In order to gain hard drive space, you may want to remove some steps from your complete analysis.

In this case, you can specify in the configuration file which step must be ERASED along the pipeline (as soon as the step following them has been correctly completed), using the key cleaner.

As an example

3=samtools sort


There only the last step result (samtools sort) will be conserved. The folders and logs of the cleaned steps are conserved, but not the resulting files.

NOTE: the last step will never be cleaned. This is true for single analyses (steps lower than 999 - script) as well as for common steps (higher than 1000 - script)

Compressing steps

BETA FEATURE: Using the compressing steps may impair the execution of some steps. Use it carefully.

In order to gain hard drive space but conserving data, you may want to compress some steps from your complete analysis.

In this case, you can specify in the configuration file which step must be COMPRESSED in tar.gz along the pipeline (as soon as the step following them has been correctly completed), using the key compress.

As an example

3=samtools sort


There only the last step result (samtools sort) will be conserved, the other being compressed in their respective tar.gz archive.

NOTE: as for the cleaner system, the last step will never be compressed. This is true for single analyses (steps lower than 999 - script) as well as for common steps (higher than 1000 - script).

BE CAREFUL: CLEANING IS OF HIGHER ORDER THAN COMPRESS. If the same step is required to be cleaned AND compressed, it will be only cleaned!

Schedulers and parallel jobs

TOGGLe is scheduler-aware (SGE, MPRUN, Slurm and LSF on v0.3+), and is able to decide by itself to launch on such a system. However, the jobs will be launched in parallel only if the software.config file contains informations for scheduling, i.e. the queue name, number of core/threads per jobs, etc…

For SGE, configuration can be provided such as:

-q myqueue.q
-pe ompi 2
-b Y

If the software.config file contains those SGE/Slurm/MPRUN/LSF informations AND the machine is SGE/Slurm/MPRUN/LSF capable, the and the scripts will be launched as parallel jobs.

Moreover, in parallel as in linear mode, an error in one individual will not stop the whole process. This individual will be marked as error in the error log, but the others will continue.

Providing a specific environment variables

If you need to provide specific ENVIRONMENT variables in your jobs (e.g. export or module load), you can provide the $env key in your software.config file

module load perl5/5.24.0
export PATH=$PATH:/my/other/PATH
export PERL5LIB=$PERL5LIB/my/new/path

Transferring files by scp

If you need to transfer the files to a temporary disk (e.g. /scratch) using scp, you can provide a $scp key followed by the temporary directory name in your configuration file


Generic Command

BETA FEATURE: if you need to launch a tool not yet integrated in TOGGLe, you can provide a Generic command using the following syntax (example of simple bash command to count SNP in a VCF file)


grep -v "#" FILEIN | wc -l > FILEOUT

The FILEIN and FILEOUT terms must be written such as, and will be automatically replaced by TOGGLe. Here in the example, the FILEIN will be modified as the previous VCF file, and FILEOUT as the output file.

BE CAREFUL: NO FORMAT CHECK will be performed, as well as no sanity control for the following step. This option is not recommanded to non-expert users.

Output and Error Logs

TOGGLe will generate two main types of logs, the .o for normal output and the .e for the errors and warnings (these last ones are normally empty files). Each level of TOGGLe will generate this pair of log:

  • TOGGLE_date_log.o/.e logs represent the general output for the complete analysis ( logs). They are located at the root of the output directory.

  • SampleName_log.o/.e logs represent the local output for sub analysis ( and logs). They are located in their respective subdirectories in the output folder.

Drawing the pipeline

If Graphviz is installed on the running server, the script will also generate a graphical view of the pipeline in the output folder. If Graphviz is not installed, a .dot file is nevertheless created, allowing the user to create a graphical view on another machine having Graphviz.

Report Option

If the -report option is set, TOGGLe will perform a report in PDF providing all the informations about the execution of the pipeline:

  • Command line provided
  • Software version
  • Basic stats about input files
  • Basic stats about the mapping, VCF, and BAM files

If you want to use this option, the system must have access to GraphViz and LaTEX distribution (TexLive recommanded)

Process_Radstacks step

If process_radstacks from stacks is used in your pipeline (for demultiplexing), you have to put it as the first step or TOGGLe will return an error and stop. After demultiplexing, TOGGLe will start from step 2 to treat all demultiplexed individuals.