Entering edit mode
8.6 years ago
komal.rathi
★
4.1k
Hi,
I am trying to apply voom to expected counts from RSEM FPKM (I don't have raw counts). My next step is to compute a differential test using limma.
# matrix I am trying to apply voom to has 1 row and 8021 columns:
> dim(tmp)
[1] 1 8021
# first five columns
> tmp[,1:5]
SRR1068687 SRR1068788 SRR1068808 SRR1068832 SRR1068855
ENSG00000149294.16 2528.61 756.53 36 158.95 1652.77
# no NAs in the matrix
> whichis.na(tmp))
integer(0)
# this is my design. shows that I have replicates.
> rbind(head(design),tail(design))
studyGTEx studyTARGET
SRR1068687 1 0
SRR1068788 1 0
SRR1068808 1 0
SRR1068832 1 0
SRR1068855 1 0
SRR1068880 1 0
f3b75630.42da.4b66.96c5.94e9a8142261 0 1
f62f350c.f8b5.4715.ace1.387f7e59cb91 0 1
f637ca92.407d.432e.ba08.2bebe05b96f4 0 1
f835cce2.1ed3.4c59.95a2.4bfef161082b 0 1
fb8c3046.eb74.43ff.ae1d.327453221555 0 1
fdee8bef.5ce2.4ca6.b6b9.3423219a1ea4 0 1
# I get an error when I try to apply voom:
> voom(tmp, design = design)
Error in approxfun(l, rule = 2) :
need at least two non-NA values to interpolate
I searched for the error and I found that it is usually encountered when you have no replicates. However in my case I have enough replicates - studyGTEx has about 7000 and the remaining are studyTARGET. What could be the problem?
NOTE: I have already tried adding more rows (to make nrow(tmp)>1
) and I still get the same error.
Crossposted: https://support.bioconductor.org/p/86934/