How do I extract read counts from DESeq2
1
1
Entering edit mode
hechtp ▴ 10
@hechtp-9860
Last seen 8.7 years ago

Hello,

I have a RNA-seq dataset in which I am adjusting for subject age (design=~age+condition).

I am wondering if DESeq2 generates a table of normalized reads after adjusting for the covariate, "age".  If so, how can I extract it?

I ask because I would like to extract the normalized reads (adjusted for the covariate) that I can use to create a gene co-expression network.
 

deseq2 counts covariates • 10k views
ADD COMMENT
1
Entering edit mode
vsd <- varianceStabilizingTransformation(dds)

You can save them:

write.table(assay(vsd), file="varianceStabilizingTransformation.txt", sep="\t")
ADD REPLY
2
Entering edit mode
@mikelove
Last seen 23 hours ago
United States

Similar to g.atla's answer, you can perform one of DESeq2's transformations (see vignette) and then, to adjust for a covariate, you can use limma's removeBatchEffect() function on the transformed values. Note that the transformed values will be on the log2 scale.

# you can use VST, rlog, or normTransform() function in DESeq2
# to create a DESeqTransform object. then:
mat <- assay(vsd) 
library(limma)
mat2 <- removeBatchEffect(mat, vsd$age.binned)

What's this age.binned I use in the last line? Read the DESeq2 FAQ about using age as a continuous covariate. In short, I do not recommend adjusting for age as a continuous covariate, and offer an alternative by creating a new variable which bins age into, say, 5 groups using the cut() function.

vsd$age.binned <- cut(vsd$age, 5)
ADD COMMENT
0
Entering edit mode

That is perfect. Thank you both for your time!

ADD REPLY

Login before adding your answer.

Traffic: 758 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