Pisces Quick Start 5.3.0

Please cite the Pisces publication when using results from this software

Pisces: An Accurate and Versatile Variant Caller for Somatic and Germline Next-Generation Sequencing Data

Introduction

Data used in paper

https://basespace.illumina.com/s/6vuFN2Cn0Ems

Suggested configuration

Somatic and germline Pisces settings below were used for the Amplicon data in the paper above. The paper results are from Pisces v5.2.5. The ultra low freq command is typically used with a UMI-collapsed BAM, and is included with the caveat that although this will find your variants, it will also call a lot of noise, so further filtering is suggested.

• Somatic: -bam {Bam} -CallMNVs false -g {genome folder} -gVCF false -i {interval file} -OutFolder {outfolder}

• Germline: -bam {Bam} -CallMNVs false -crushvcf true -g {genome folder} -gVCF false -i {interval file} -ploidy diploid -OutFolder {outfolder}

• Ultra low freq: -bam {Bam} -g {genome folder} -OutFolder {outfolder} -MinVF 0.0005 -SSFilter false -MinBQ 65 -MaxVQ 100 -MinDepthFilter 500 -MinVQ 0 -VQFilter 20 -ReportNoCalls True -CallMNVs False -MinDepth 5 -threadbychr true

• High Speed: (To get results on a exome / WGS bam fast as possible) -bam {Bam} -CallMNVs false -g {genome folder} -gVCF false -OutFolder {outfolder} -ThreadByChr True -FilterDuplicates true -EnableSingleStrandFilter true

Please note that exact syntax might change, version to version. The version-specific commands are directly available from the command line help, or from the version-specific wiki page ie, https://github.com/tamsen/Pisces/wiki/Pisces-5.2.9-Supported-Options.

For versions before 5.2.9, the RMxNFilter frequency threshold value was set at 20%, not 35%. So, you would have to append " -RMxNFilter 5,9,0.35" to prior versions for similar performance.

Docker Files

https://github.com/tamsen/Pisces/tree/master/docker