read counts of paired end reads for DESeq2 analysis
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tszn1984 • 0
@tszn1984-10590
Last seen 8.6 years ago

I got a  list of paired end bam files, and want to do DESeq2 analysis. The first thing is to count the reads at gene level. I tried feature counts. However, it requires reorder of the bam files if I specified paired end is True, which takes infinite time for my huge dataset. I also tried summarizeOverlaps, which also takes very long time. 

Is there any tools could quickly count the paired end reads for DESeq2 at gene level? Thanks.

deseq featurecounts paired • 2.1k views
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Are you using the latest version of featureCounts (part of Rsubread package)? Running time for re-ordering read pairs has been significantly reduced in recent releases of featureCounts. FeatureCounts now only needs to re-order a small portion of read pairs instead of re-ordering all read pairs as it performed in older version. It now takes featureCounts less than one minute to count 30 million read pairs with 4 threads (this includes the read re-ordering time).

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@steve-lianoglou-2771
Last seen 22 months ago
United States

If you're looking for speed, you might try using kallisto or salmon to quantitate expression from your data, then use the tximport package to roll the transcript-level estimates up into gene level estimates so you can feed those numbers into DESeq2 (or edgeR (or voom)).

Note that, technically, you will have to be running the development version of Bioconductor to get tximport ...

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Yes, I'd try that, and add that tximport will be in the release branch in about a week (~May 4).

Also, for featureCounts, you could name-sort your BAM files using samtools with multiple threads...

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