Changes between Version 37 and Version 38 of SOPs/atac_Seq
- Timestamp:
- 03/16/21 11:57:52 (4 years ago)
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SOPs/atac_Seq
v37 v38 136 136 Additional 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. 137 137 * 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. 138 139 * another approach is to convert the bam file to bed (using bedtools), and use the options, -f BED --shift 100 --extsize 200 139 140