DESeq2: seeking advice regarding before/after mutliple treatements + extra factor
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Last seen 2.4 years ago


I am trying to analyze my 16S data using DESeq2, and I would like some advice on the correct formula I should use.

I have before/after data for two different treatments; also, I have 2 age classes. This is a sample of the table:

Patient Time    Treatment   AgeClass
p1      Before       A           C
p1      After        A           C
p2      Before       A           M
p2      After        A           M
p3      Before       B           C
p3      After        B           C
p4      Before       B           M
p4      After        B           M

As you can see, Treatment has two levels (A and B). I want to look at the effect of the two treatments, but also taking into account the age class of the patient. I was wondering if the following formula is correct:

~AgeClass + Treatment + Treatment:Patient + Treatment:Time

Should I make any change to the formula?

Thank you for your kind attention, Max

deseq2 16s microbiome • 314 views
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Last seen 19 hours ago
United States

My first note is that I'm not sure DESeq2 is appropriate for microbiome datasets necessarily. There was a recent review paper posted to bioRxiv so maybe you can read that to determine if it's appropriate for your data.

Otherwise, this is a common question on Bioconductor. If you control for patient, you should not also add patient characteristics to the design. This is unnecessary and will produce a linear dependence in the design.

Entering edit mode

Thank you for the kind answer, and for the link to the paper, I appreciate it. I will read it and review my pipeline accordingly. I am sorry if my question has been asked repeatedly, I read a few topics and still I was not sure about my code.

Regarding the variables, for what I understand then, it should be either

~Treatment + Treatment:Patient + Treatment:Time


~AgeClass + Treatment + Treatment:Time

but never "AgeClass" and "Patient" together.

Thanks again for taking the time to answering me.




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