use tximport's countsFromAbundance when using DESeq2
1
0
Entering edit mode
ginlucks • 0
@ginlucks-15159
Last seen 3.7 years ago
Italy

Hi all,

I was wondering if using tximport to import salmon qf files I have to use the countsFromAbundance option (it is meant to score counts using abundance estimates scaled up to library size) since DESeq2 uses un-normalized data and since by the "counts" function it is possible to get normalized counts.

deseq2 tximport normalization salmon rnaseq • 2.7k views
ADD COMMENT
0
Entering edit mode
@mikelove
Last seen 9 hours ago
United States

Yes, you can use countsFromAbundance="lengthScaledTPM" or "scaledTPM" as input to DESeqDataSetFromMatrix. 

I prefer the DESeqDataSetFromTximport approach myself (which calculates an offset matrix for the user, and uses the estimated counts directly), but it's not a big difference. The counts + offset approach is the typical way to deal with biases in a GLM setting.

The countsFromAbundance approach instead scales the counts up and down per gene to account for any change in gene length due to differential isoform usage. Because this event is not that common or severe -- globally -- it doesn't end up scaling the counts that much for most genes. But you should still use one or the other approach (counts + offset or countsFromAbundance) to protect against the potential bias if there were differential isoform usage among isoforms of substantially different length across samples for a particular gene (e.g. as described by Trapnell 2013).

ADD COMMENT
0
Entering edit mode

Thanks for answer

 

ADD REPLY

Login before adding your answer.

Traffic: 698 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6