CUT&RUN (and CUT&TAG) Workflow

public public 1yr ago Version: v0.3 0 bookmarks

Inputs dataset

  • The workflow needs a single input which is a list of dataset pairs of fastqsanger.

Inputs values

  • adapter sequences: this depends on the library preparation. Usually CUT&RUN is Truseq and CUT&TAG is Nextera. If you don't know, use FastQC to determine if it is Truseq or Nextera

  • reference_genome: this field will be adapted to the genomes available for bowtie2

  • effective_genome_size: this is used by macs2 and may be entered manually (indications are provided for heavily used genomes)

  • normalize_profile: Whether you want to have a profile normalized as Signal to Million Reads.

Processing

  • The workflow will remove illumina adapters and low quality bases and filter out any read smaller than 15bp

  • The filtered reads are mapped with bowtie2 allowing dovetail and fragment length up to 1kb

  • The BAM is filtered to keep only MAPQ30 and concordant pairs

  • The PCR duplicates are removed with Picard

  • The BAM is converted to BED to enable macs2 to take both pairs into account

  • The peaks are called with macs2 which at the same time generates a coverage file (normalized or not).

  • The coverage file is converted to bigwig

  • A multiQC is run to have an overview of the QC

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Free

Created: 1yr ago
Updated: 1yr ago
Maitainers: public
URL: https://github.com/iwc-workflows/cutandrun
Name: cutandrun-main
Version: v0.3
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Copyright: Public Domain
License: MIT License
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