[BioC] Problem with topTable function (after Bioconductor update)
Christian De Santis
christian.desantis at stir.ac.uk
Wed Oct 30 19:37:47 CET 2013
Hi Jim,
Thank for the detailed explanation and yes, there was some humour together with some truth! :-)
It really drove me crazy as it was working up until yesterday. That's why I thought it must have come with the Bioconductor update.
I tried what you said and it worked just fine!!! Just a note for someone else might have the same problem and read this post, there was a misspelling in your code (P.Value instead of P.value).
Thanks so much for taking the time to answer and rescue me. I really appreciated it!
Regards,
Christian
-----Original Message-----
From: James W. MacDonald [mailto:jmacdon at uw.edu]
Sent: 30 October 2013 18:08
To: Christian De Santis
Cc: bioconductor at r-project.org
Subject: Re: [BioC] Problem with topTable function (after Bioconductor update)
Hi Christian,
On Wednesday, October 30, 2013 12:23:46 PM, Christian De Santis wrote:
> Hi to everybody,
>
> I hope someone can help me on this as I am losing my health.
Not sure if you were intending humor, but this cracked me up!
>
> I was re running my script today which worked fine till last time I did it. When I run the topTable function on more than one coefficient it gives me the following error.
>
>> x.contrast.REG <- topTable(fit.contrast, adjust.method="none",
>> coef=c(1:4), number=20000, p.value = 0.05)
> Error in `row.names<-.data.frame`(`*tmp*`, value = value) :
> duplicate 'row.names' are not allowed In addition: Warning message:
> non-unique values when setting 'row.names':
>
> I have tried to run the same script for each coefficient separately and it works fine. I can't explain the error since if there were duplicated row.names then I should get the same error when doing the analysis on individual contrasts since I run topTable on the fit object.
>
> The only thing I have changed was the fact that this morning I updated Bioconductor to the latest release.
>
> I really hope someone can help me understand.
There appears to be a bug in the function topTableF.
When you run topTable() on multiple coefficients, it actually uses topTableF(). In that function there is a bit of code where the data are subsetted according to the p.value and lfc values you submitted:
if (lfc > 0 || p.value < 1) {
if (lfc > 0)
big <- rowSums(abs(M) > lfc, na.rm = TRUE) > 0
else big <- TRUE
if (p.value < 1) {
sig <- adj.P.Value <= p.value
sig[is.na(sig)] <- FALSE
}
else sig <- TRUE
keep <- big & sig
if (!all(keep)) {
M <- M[keep, , drop = FALSE]
rn <- rn[keep]
Amean <- Amean[keep]
Fstat <- Fstat[keep]
Fp <- p.value[keep]
genelist <- genelist[keep, , drop = FALSE]
adj.P.Value <- adj.P.Value[keep]
}
So now all the data have been subsetted (in your case) to just those genes with an unadjusted p-value < 0.05. And presumably that is fewer than the 20000 genes that you started with. But then there is some sorting of the resulting data:
if (sort.by == "F")
o <- order(Fp, decreasing = FALSE)[1:number]
else o <- 1:number
if (is.null(genelist))
tab <- data.frame(M[o, , drop = FALSE])
else tab <- data.frame(genelist[o, , drop = FALSE], M[o,
, drop = FALSE])
tab$AveExpr = fit$Amean[o]
tab <- data.frame(tab, F = Fstat[o], P.Value = Fp[o], adj.P.Val =
adj.P.Value[o])
rownames(tab) <- rn[o]
The problem here is that 1:number is _longer_ than Fp, which has been subsetted already, based on the p-value criterion. So the vector 'o'
will have a bunch of NAs on the end. As an example:
(1:5)[1:8]
[1] 1 2 3 4 5 NA NA NA
And when you get to the last line there, where the rownames are set, since there are multiple NAs, you will have repeated rownames, which R won't allow.
In the interest of your health, you can get around this for now by doing
num <- sum(topTable(fit.contrast, adjust.method="none", coef=c(1:4), number=20000)$P.value < 0.05)
and then
x.contrast.REG <- topTable(fit.contrast, adjust.method="none", coef=c(1:4), number=num, p.value = 0.05)
Best,
Jim
>
> Regards,
> Christian De Santis
>
>> sessionInfo()
> R version 3.0.1 (2013-05-16)
> Platform: i386-w64-mingw32/i386 (32-bit)
>
> locale:
> [1] LC_COLLATE=English_United Kingdom.1252 LC_CTYPE=English_United Kingdom.1252 LC_MONETARY=English_United Kingdom.1252
> [4] LC_NUMERIC=C LC_TIME=English_United Kingdom.1252
>
> attached base packages:
> [1] stats graphics grDevices utils datasets methods base
>
> other attached packages:
> [1] reshape2_1.2.2 ellipse_0.3-8 genefilter_1.44.0 limma_3.18.1 BiocInstaller_1.12.0
>
> loaded via a namespace (and not attached):
> [1] annotate_1.40.0 AnnotationDbi_1.24.0 Biobase_2.22.0 BiocGenerics_0.8.0 DBI_0.2-7
> [6] IRanges_1.20.3 parallel_3.0.1 plyr_1.8 RSQLite_0.11.4 splines_3.0.1
> [11] stats4_3.0.1 stringr_0.6.2 survival_2.37-4 tools_3.0.1 XML_3.98-1.1
> [16] xtable_1.7-1
>
>
>
>
--
James W. MacDonald, M.S.
Biostatistician
University of Washington
Environmental and Occupational Health Sciences
4225 Roosevelt Way NE, # 100
Seattle WA 98105-6099
--
The University of Stirling has been ranked in the top 12 of UK universities for graduate employment*.
94% of our 2012 graduates were in work and/or further study within six months of graduation.
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