Flexiprep

Introduction

Flexiprep is a quality control pipeline. This pipeline checks for possible barcode contamination, clips reads, trims reads and runs FASTQC. Adapter clipping is performed by Cutadapt. For quality trimming we use Sickle. Flexiprep only works on .fastq files.

Example

To get the help menu:

biopet pipeline Flexiprep -h

Arguments for Flexiprep:
 -R1,--input_r1 <input_r1>             R1 fastq file (gzipped allowed)
 -R2,--input_r2 <input_r2>             R2 fastq file (gzipped allowed)
 -sample,--sampleid <sampleid>         Sample ID
 -library,--libid <libid>              Library ID
 -config,--config_file <config_file>   JSON config file(s)
 -DSC,--disablescatter                 Disable all scatters

Note that the pipeline also works on unpaired reads where one should only provide R1.

To start the pipeline (remove -run for a dry run):

biopet pipeline Flexiprep -run -outDir myDir \
-R1 myFirstReadPair -R2 mySecondReadPair -sample mySampleName \
-library myLibname -config mySettings.json

Configuration and flags

For technical reasons, single sample pipelines, such as this pipeline do not take a sample config. Input files are in stead given on the command line as a flag.

Command line flags for Flexiprep are:

Flag (short)	Flag (long)	Type	Function
-R1	--inputR1	Path (required)	Path to input fastq file
-R2	--inputR2	Path (optional)	Path to second read pair fastq file.
-sample	--sampleid	String (required)	Name of sample
-library	--libid	String (required)	Name of library

If -R2 is given, the pipeline will assume a paired-end setup.

Sample input extensions

Please refer to our mapping pipeline for information about how the input samples should be handled.

Config

All other values should be provided in the config. Specific config values towards the mapping pipeline are:

Name	Type	Function
skip_trim	Boolean	Default false, if true the trimming step is skipped
skip_clip	Boolean	Default false, if true the clipping step is skipped

Result files

The results from this pipeline will be a fastq file. The pipeline also outputs 2 Fastqc runs one before and one after quality control.

Example output

.
├── mySample_01.qc.summary.json
├── mySample_01.qc.summary.json.out
├── mySample_01.R1.contams.txt
├── mySample_01.R1.fastqc
│   ├── mySample_01.R1_fastqc
│   │   ├── fastqc_data.txt
│   │   ├── fastqc_report.html
│   │   ├── Icons
│   │   │   ├── error.png
│   │   │   ├── fastqc_icon.png
│   │   │   ├── tick.png
│   │   │   └── warning.png
│   │   ├── Images
│   │   │   └── warning.png
│   │   ├── Images
│   │   │   ├── duplication_levels.png
│   │   │   ├── kmer_profiles.png
│   │   │   ├── per_base_gc_content.png
│   │   │   ├── per_base_n_content.png
│   │   │   ├── per_base_quality.png
│   │   │   ├── per_base_sequence_content.png
│   │   │   ├── per_sequence_gc_content.png
│   │   │   ├── per_sequence_quality.png
│   │   │   └── sequence_length_distribution.png
│   │   └── summary.txt
│   └── mySample_01.R1.qc_fastqc.zip
├── mySample_01.R1.qc.fastq.gz
├── mySample_01.R1.qc.fastq.gz.md5
├── mySample_01.R2.contams.txt
├── mySample_01.R2.fastqc
│   ├── mySample_01.R2_fastqc
│   │   ├── fastqc_data.txt
│   │   ├── fastqc_report.html
│   │   ├── Icons
│   │   │   ├── error.png
│   │   │   ├── fastqc_icon.png
│   │   │   ├── tick.png
│   │   │   └── warning.png
│   │   ├── Images
│   │   │   ├── duplication_levels.png
│   │   │   ├── kmer_profiles.png
│   │   │   ├── per_base_gc_content.png
│   │   │   ├── per_base_n_content.png
│   │   │   ├── per_base_quality.png
│   │   │   ├── per_base_sequence_content.png
│   │   │   ├── per_sequence_gc_content.png
│   │   │   ├── per_sequence_quality.png
│   │   │   └── sequence_length_distribution.png
│   │   └── summary.txt
│   └── mySample_01.R2_fastqc.zip
├── mySample_01.R2.fastq.md5
├── mySample_01.R2.qc.fastqc
│   ├── mySample_01.R2.qc_fastqc
│   │   ├── fastqc_data.txt
│   │   ├── fastqc_report.html
│   │   ├── Icons
│   │   │   ├── error.png
│   │   │   ├── fastqc_icon.png
│   │   │   ├── tick.png
│   │   │   └── warning.png
│   │   ├── Images
│   │   │   ├── duplication_levels.png
│   │   │   ├── kmer_profiles.png
│   │   │   ├── per_base_gc_content.png
│   │   │   ├── per_base_n_content.png
│   │   │   ├── per_base_quality.png
│   │   │   ├── per_base_sequence_content.png
│   │   │   ├── per_sequence_gc_content.png
│   │   │   ├── per_sequence_quality.png
│   │   │   └── sequence_length_distribution.png
│   │   └── summary.txt
│   └── mySample_01.R2.qc_fastqc.zip
├── mySample_01.R2.qc.fastq.gz
├── mySample_01.R2.qc.fastq.gz.md5
└── report

Getting Help

If you have any questions on running Flexiprep, suggestions on how to improve the overall flow, or requests for your favorite Quality Control (QC) related program to be added, feel free to post an issue to our issue tracker at GitHub. Or contact us directly via: SASC email