DESeq2 Control for differences between patients
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gkuffel ▴ 10
@gkuffel-12871
Last seen 5.9 years ago
Loyola University

Hi,

I have a relatively simple RNA-Seq experiment. I have 10 samples in my control group and 10 samples in my treatment group. Each sample is from a different patient. How can I test for differences between control and treatment groups but control for the natural differences we might see between patients. Your help is greatly appreciated.

deseq2 • 968 views
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swbarnes2 ★ 1.4k
@swbarnes2-14086
Last seen 2 hours ago
San Diego

You have 10 control samples and 10 treatment samples.  If you wanted to control for sex, or age (maybe by splitting the samples into a few age groupings) you could tell it that information, and make use of that, but it's already going to treat your samples as if they were biological replicates.

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Thank you for the response. So I do not see any differential expression when using design =~ condition and p-adj < 0.1. Is there anything else I can modify or try? Also, if I look at p values instead of p-adj then I see some differential expression (about 1000 genes).

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Unadjusted p-values are not the right thing to be looking at.  Stick with adjusted p-values.  As always, XKCD explains why:  https://xkcd.com/882/

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Take a look at the MA plot (see vignette). This will give you a senses of the differences across treatment. You can also look at the plotCounts() function to see why the top gene doesn't have an adjusted p-value less than 0.1. This typically means small differences across group relative to the variance within group.

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