[BioC] DESeq/DESeq normalization on different experiments
Ryan
rct at thompsonclan.org
Thu May 22 17:06:34 CEST 2014
Hi Gilgi,
If you are not going to test for differential expression between
experiments, then there is no purpose in normalizing them together. The
more worrying problem with analyzing all your experiments as a single
data set is that a single dispersion value will be estimated for each
gene across all experiments. This is only ok if you believe that every
gene has equal biological variability in all your experiments, which is
unlikely to be the case.
-Ryan
On Thu May 22 01:26:55 2014, Gilgi [guest] wrote:
> Hello,
>
> I have ~100 RNA seq-samples from the same organism, but they include different experiments (each experiment of 6-16 samples). I would like to put all together into our RNA seq pipeline, that includes mapping, htseq count and DESeq. Differential expression comparisons will be done of course only between samples of a single experiment (due to possible batch effects). However, will it be a good idea to normalize all samples together?
> The assumption behind the normalization is that most genes are not differentially expressed, but between different experiments the variability might be higher. Therefore I wanted to ask your opinion on doing the normalization on such a large combined data-set.
>
> Thank you,
> Gilgi
>
> -- output of sessionInfo():
>
> none
>
> --
> Sent via the guest posting facility at bioconductor.org.
>
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