Running DESeq2 without experimental conditions
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3.5 years ago
grsyhhsb ▴ 10

Hi all! I tried running DESeq2 differential expression analysis on some RNA seq data without adding experimental conditions and DESeq2 actually outputs the results including log2 fold change and p-vals. Does anyone know what the results mean since there aren't any groups to compare with? Thank you!

RNA-seq DESeq2 differential-expression • 822 views
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Can you show code? It is to my knowledge not possible to run DESeq2 without a design, the only thing I could imagine is to run as intercept-only ~1 but that makes no sense in a DE setting, see https://support.bioconductor.org/p/79209/

Or do you mean running results() without explicit name or contrast?

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Hello! Thanks for the reply. I did set it to run as intercept only, which I realized doesn't really mean anything. I was hoping to find genes that have the highest variability across samples. Upon doing some research, I think I should do Median Absolute Deviation analysis.

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Do you have experimental replicates? Consider the LRT test in DESeq2 to find any differences between the groups. MAD does not mean a lot if you do not properly model variance as a function of the mean because variance is not equal across genes with different expression levels. The concepts of heteroscedasticity and overdispersion kick in here, which is why you should use expert software rather than custom approaches.

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