ATAC-Seq and featureCounts

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3 months ago

qudrat.nii ▴ 10

Hello,

I have ATAC-Seq bam files and I want to know the quality of files, so I run featureCounts. I found the following result...

Successfully assigned alignments : 54255620 (40.3%)

Is it OK or what should be the percentage of assigned alignment?

ATAC-Seq featureCounts • 637 views

ADD COMMENT • link updated 3 months ago by ATpoint 82k • written 3 months ago by qudrat.nii ▴ 10

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If this is the percentage of reads overlapping callable peaks then this actually pretty good. Some context on commands and what you use as annotations here to count would be helpful though.

ADD REPLY • link 3 months ago by ATpoint 82k

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I use following command:

featureCounts - p -a peak.saf -F SAF -o readCountInPeaks.txt ${sample}.bam

ADD REPLY • link updated 3 months ago by GenoMax 141k • written 3 months ago by qudrat.nii ▴ 10

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If you are using -p for paired end reads then you will want to use following as well to count fragments.

--countReadPairs

ADD REPLY • link 3 months ago by GenoMax 141k

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As said, if this is reads in peaks (FRiPs) then this is fine.

ADD REPLY • link 3 months ago by ATpoint 82k

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Sort of impossible to answer without more specific information. What organism? What annotation? Has anyone ever done ATAC Seq in this organism before? Do you have a positive control experiment (even in another organism to control for your analysis pipeline) that you can compare the number to?

ADD REPLY • link 3 months ago by seidel 11k

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