[BioC] error in mas5calls() for soybean chip

Jenny Drnevich drnevich at uiuc.edu
Wed Apr 19 19:07:13 CEST 2006


Hi Jim and Ken and the rest of the list,

Thanks for the debugging suggestions. The problem is indeed with "probe 
sets" that only have one probe pair; if the PM > MM, then 'DetectionPValue' 
returns 0, but it PM <= MM then 'NaN' is returned. I was able to get 
'mas5calls.AffyBatch' to work by simply adding this one command before the 
P/M/A calls were made:

p[sapply(p,is.na)] <- 1


I'm not sure what's the best way to call P/M/A when there is only one probe 
pair in a probe set, but perhaps a warning/output could be generated along 
the lines of "One or more of the probe sets has only one probe pair. Use 
'oneProbePair()' to find out which ones". [code is below for a simple 
function I wrote to get the probe set names]. Or would a warning cause more 
problems from unnecessary panic? I was surprised that there were 92 "probe 
sets" on the soybean chip with only one probe pair? I wonder how many other 
chips actually have single probe pair "probe sets" - as long as PM>MM for 
all the arrays, mas5calls() would work and you'd never know...

Cheers,
Jenny

oneProbePair <- function(object) {
     x <- probeNames(object)
     dups <- unique(x[duplicated(x)])
     singles <- setdiff(unique(x),dups)
     return(singles)
     }


At 01:49 PM 4/18/2006, James W. MacDonald wrote:
>Hi Jenny,
>
>Jenny Drnevich wrote:
> > Hi all,
> >
> > I've run into some sort of error or bug when trying to get Affy's
> > present/marginal/absent calls for  Affy's soybean array. I've traced the
> > error to this part of mas5calls.AffyBatch():
> >
> > calls <- sapply(p, function(y) {
> >          if (y < alpha1) {
> >
> >
> > According to Benjamin Otto's reponse
> > (https://stat.ethz.ch/pipermail/bioconductor/2006-January/011568.html) to
> > the exact same error message for  HU6800 arrays, specifying alpha1= and
> > alpha2= in the call to mas5calls() should work, but it doesn't for soybean
> > arrays (all codes & output at the bottom). I'm actually not surprised that
> > it doesn't work, because these are the defaults in the internal function
> > mas5calls.AffyBatch() that's called.  Is problem this related to similar
> > problems with the qc() function from simpleaffy not working for some array
> > types? I'll try the solution suggested in
> > https://stat.ethz.ch/pipermail/bioconductor/2006-April/012655.html, but
> > mas5calls is part of the affy package, and I didn't see where functions
> > from simpleaffy were called.
>
>This problem isn't related to anything in simpleaffy. I am betting that
>there are some NAs in the p list that is returned from the call to
>'DetectionPValue' (immediately preceding the code snippet you give
>above). Without actually having any soybean arrays to play with, I
>cannot confirm this, but you could by stepping through the code using
>
>debug(mas5calls.AffyBatch)
>mas5calls(rawdata[,1:5])
>
>when you get to the point that the 'p' list is made, you can dump it
>into your .GlobalEnv using
>
>p <<- p
>
>and then quit the debugger (using Q). You could then do something like
>
>sum(sapply(p, is.na))
>
>to see how many (or if) there are any NAs in there. You could also
>figure out which probesets are returning the NAs, and take a look at
>them using the PM() accessor to see if you can tell what the problem
>might be.
>
>Any information you might be able to give could be helpful in debugging
>this error.
>
>Best,
>
>Jim
>
>
> >
> >
> > Thanks,
> > Jenny
> >
> >  > rawdata
> > AffyBatch object
> > size of arrays=1164x1164 features (317565 kb)
> > cdf=Soybean (61170 affyids)
> > number of samples=30
> > number of genes=61170
> > annotation=soybean
> >
> >  > PMAcalls <- mas5calls(rawdata[,1:5])
> > Getting probe level data...
> > Computing p-values
> > Making P/M/A Calls
> > Error in if (y < alpha1) { : missing value where TRUE/FALSE needed
> >
> >  > PMAcalls <- mas5calls(rawdata[,1:5],alpha1=0.04,alpha2=0.06)
> > Getting probe level data...
> > Computing p-values
> > Making P/M/A Calls
> > Error in if (y < alpha1) { : missing value where TRUE/FALSE needed
> >
> >
> >  > sessionInfo()
> > R version 2.2.1, 2005-12-20, i386-pc-mingw32
> >
> > attached base packages:
> > [1] "splines"   "tools"     "methods"   "stats"     "graphics"  "grDevices"
> > [7] "utils"     "datasets"  "base"
> >
> > other attached packages:
> >     soybeancdf         made4 scatterplot3d          ade4  affyQCReport
> >       "1.10.0"       "1.4.0"      "0.3-24"       "1.4-0"       "1.8.0"
> >     simpleaffy    genefilter      survival    reposTools       affyPLM
> >        "2.4.2"       "1.8.0"        "2.20"       "1.8.0"       "1.6.0"
> >       affydata         limma         gcrma   matchprobes          affy
> >        "1.6.0"       "2.4.7"       "2.2.0"       "1.2.1"       "1.8.1"
> >        Biobase       RWinEdt
> >        "1.8.0"       "1.7-4"
> >
> >
> >
> > Jenny Drnevich, Ph.D.
> >
> > Functional Genomics Bioinformatics Specialist
> > W.M. Keck Center for Comparative and Functional Genomics
> > Roy J. Carver Biotechnology Center
> > University of Illinois, Urbana-Champaign
> >
> > 330 ERML
> > 1201 W. Gregory Dr.
> > Urbana, IL 61801
> > USA
> >
> > ph: 217-244-7355
> > fax: 217-265-5066
> > e-mail: drnevich at uiuc.edu
> >
> > _______________________________________________
> > Bioconductor mailing list
> > Bioconductor at stat.math.ethz.ch
> > https://stat.ethz.ch/mailman/listinfo/bioconductor
> > Search the archives: 
> http://news.gmane.org/gmane.science.biology.informatics.conductor
>
>
>--
>James W. MacDonald, M.S.
>Biostatistician
>Affymetrix and cDNA Microarray Core
>University of Michigan Cancer Center
>1500 E. Medical Center Drive
>7410 CCGC
>Ann Arbor MI 48109
>734-647-5623
>
>
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Jenny Drnevich, Ph.D.

Functional Genomics Bioinformatics Specialist
W.M. Keck Center for Comparative and Functional Genomics
Roy J. Carver Biotechnology Center
University of Illinois, Urbana-Champaign

330 ERML
1201 W. Gregory Dr.
Urbana, IL 61801
USA

ph: 217-244-7355
fax: 217-265-5066
e-mail: drnevich at uiuc.edu



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