Hi @Alok_Dubey
Replicates will be required to use this tool and the other Bioconductor DE tools.
FAQ Extended Help for Differential Expression Analysis Tools
And, here is a summary from another person on our team, who captures the situation well, so am just quoting.
Yes, that is the problem.
See also DESeq2 without biol replicates - SEQanswers where Simon Anders and Michael Love chime in, the original authors of DESeq and DESeq2.
I think the best you can do is to normalize the counts and look at the fold changes, but none of that is really statistically sound.
Beyond that,
for the immediate error (“Tool Standard Error” in the 4th screenshot) is complaining about not understanding how to parse the “factor name”. That could be due to the term starting with a number, or from it including a space. The tool form attempts to fix the formatting to be “R friendly” but can’t always do that perfectly.
Adjusting all labels on the form can address that part. “all oneWord, no spaces, alphanumeric characters, not starting with a number, and optional underscores but not starting with one.”
If you do that ^^, and rerun, you’ll probably get the “no replicates found!” type of error.
Thanks for sharing all the details! Super helpful