Question: nomarlizing sequencing depth when use macs2 bdgbroadpeaks
0
gravatar for langya
2.4 years ago by
langya70
langya70 wrote:

Hey

I was wondering when I use macs2 bdgbroadpeaks to call broad peaks on bedgraph file (which is converted from bigwig file that is converted from bam file by our collaborator, don't ask me why they did this and they couldnt find the original bam file), how do I normalize the sequencing depth by million like macs2 callpeak options. And i notice the only output from this is the bed file:

macs2 bdgbroadcall -i fileNameExample.bw.bedGraph --o-prefix fileNameExample.bw.bedGraph

sequencing chip-seq macs2 • 691 views
ADD COMMENTlink written 2.4 years ago by langya70
1

It's expecting the input file to be appropriately normalized, if I remember correctly. If you know the original depth (or can at least eye-ball it) then you could use wiggletools to scale the file until you get something reasonable.

ADD REPLYlink written 2.4 years ago by Devon Ryan96k

they couldnt find the original bam file

can they find the FASTQ files? If not, they'll be in trouble upon submission anyway as they are going to be required by most journals to submit the raw data to a repository

You'll most likely be much better off to re-do the alignment and do things properly

ADD REPLYlink written 2.4 years ago by Friederike6.1k
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