Hi,
I followed your suggestion to calculate the p-value of my microarray and ChIP-seq data:
# bound chip-seq = 5673
# down microarray = 3975
# overlap = 1156
# N = 17970
m <- matrix(c(1156,3975-1156,5673-1156,17970-3975-5673+1156), ncol=2,
dimnames=list(c("ChIP.Sig", "ChIP.NoSig"), c("RNA.Sig","RNA.NoSig")))
fisher.test(m)
# Fisher's Exact Test for Count Data
data: m
p-value = 0.0001287
alternative hypothesis: true odds ratio is not equal to 1
95 percent confidence interval:
0.7958616 0.9299321
sample estimates:
odds ratio
0.8604837
Then I wanted to check if a random subset of the same size could be significant too, the overlap of random set with binding was 683. But when I do the calculation, the p-value is much more significant (p-value < 2.2e-16, the lowest result by Fisher test in R). I don't understand why a lower overlap can be more significant....
Am I missing something? Maybe I don't really need to check with a random gene set? This is a reply for reviewers because they suggest that the binding of my TF won't be significant over a random subset, is it enough for reply the p-value I got from Fisher test?
Thanks in advance Gema

Sorry for the big letters... I don't know why they look like that...
I think that whatever the results of a hypergeometric test might be in this case, a lot of caution should be advised in interpreting that result.
it is well-known that Chip-Seq, RNA-seq and other NGS based assays of the epigenome and transcriptome measure a lot of correlated outcomes (e.g. shared epigenetic programs affecting many genes).
It is very conceivable that in an example like the one you describe, it could be a much smaller number of factors that lead you to detect 37 genes in the overlap bin. If this is the case, using a test like the hypergeometric would be anti-conservative, and a lot of readers and reviewers might distrust the result for that reason.