- The workflow needs a single input which is a list of fastqsanger files.
- adapters sequence_forward: this depends on the library preparation. 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.
- 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 with default parameters.
- The BAM is filtered to keep only MAPQ30.
- The peaks are called with MACS2 with a fixed extension of 200bp which at the same time generates a coverage file (normalized or not).
- The coverage is converted to bigwig.
- A MultiQC is run to have an overview of the QC.
- The filtered bam still has PCR duplicates which are removed by MACS2.