Changes between Version 37 and Version 38 of SOPs/atac_Seq


Ignore:
Timestamp:
03/16/21 11:57:52 (4 years ago)
Author:
thiruvil
Comment:

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  • SOPs/atac_Seq

    v37 v38  
    136136Additional notes on calling peaks using MACS: ''using MACS default options without BAMPE will not call the correct peaks''.  Alternative options/parameters in calling peaks using MACS, if BAMPE is not used are, --nomodel --shift s --extsize 2s, this can be used for single-end reads as well, e.g. --shift 100 --extsize 200 is suitable for ATAC-Seq.   
    137137    * MACS' author, T.Liu, recommends using -f BAMPE if PE reads are used [[https://github.com/taoliu/MACS/issues/331]], using BAMPE option asks MACS to pileup and calculate the extension size -  works for finding accessible regions within cut sites.  The additional parameters can also be used to look only at the //exact// cut sites by Tn5 instead of the open/accessible regions [[https://github.com/taoliu/MACS/issues/145]], if so, -f BAMPE may not be suitable.
     138      * Shifting reads, pos. strand + 4 and neg strand -5 (see recommendations below) may be needed as well to find //exact// cut sites.
    138139    * another approach is to convert the bam file to bed (using bedtools), and use the options, -f BED --shift 100 --extsize 200
    139140