Limma: NaNs in intraspotCorrelation()
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Pie Muller ▴ 110
@pie-muller-1349
Last seen 9.6 years ago
Dear Gordon and Bioconductor list members, I am using limma to analyse an unlinked two-colour array experiment consisting of 31 arrays (with 34,992 spots on each array) and 38 unique targets. I would like to separate the two channels to perform a separate channel analysis using the function lmscFit() and tried to follow the example given in the limma user's guide (see below) but found that the "intraspotCorrelation()" gives me nothing but NaNs. What could have gone wrong? There are no NaNs in either the M or A values in my MAlist object. I suspect there might something wrong with my design matrix as I noticed that by reducing the design matrix to one single factor such as "design=matrix(rep(1,62), ncol=1)" I receive a consensus correlation value back. However, there is still another warning message "reml: Max iterations exceeded in: remlscore(y, X, Z)". Does anyone have suggestions as to solve this problem? In advance, many thanks for your support! Pie My target file: > targets Array Cy3 Cy5 1 2007-06-07_B2162-09.gpr F.168.C F.72.B 2 2007-06-07_B2162-13.gpr U.48.A U.24.A 3 2007-06-07_B2162-16.gpr F.24.E U.24.C 4 2007-06-07_B2162-20.gpr F.8.C U.8.C 5 2007-06-07_B2162-22.gpr U.24.A U.8.A 6 2007-06-15_B2162-02.gpr F.48.B F.24.D 7 2007-06-15_B2162-04.gpr U.72.A U.48.A 8 2007-06-15_B2162-05.gpr F.72.B F.48.B 9 2007-06-15_B2162-07.gpr F.48.C U.24.C 10 2007-06-15_B2162-10.gpr F.168.C U.8.B 11 2007-06-15_B2162-11.gpr U.168.A U.72.A 12 2007-06-15_B2162-19.gpr F.8.A U.168.A 13 2007-06-15_B2162-23.gpr F.72.C U.72.C 14 2007-06-15_B2162-25.gpr F.24.D F.8.B 15 2007-06-15_B2162-26.gpr F.168.D U.168.C 16 2007-09-26_B2165-05.gpr I.72.A I.48.A 17 2007-09-26_B2165-08.gpr I.24.A I.8.A 18 2007-09-26_B2165-12.gpr I.8.A F.168.B 19 2007-09-26_B2165-15.gpr I.8.A I.168.B 20 2007-09-26_B2165-18.gpr I.168.A I.72.A 21 2007-09-26_B2165-22.gpr I.48.A I.24.A 22 2007-09-28_B2165-02.gpr U.168.C I.168.C 23 2007-09-28_B2165-03.gpr I.168.C F.168.C 24 2007-09-28_B2165-04.gpr I.24.B F.24.D 25 2007-09-28_B2165-11.gpr I.72.B F.72.B 26 2007-09-28_B2165-14.gpr U.8.C I.8.C 27 2007-09-28_B2165-17.gpr U.72.C I.72.C 28 2007-09-28_B2165-19.gpr I.48.B F.48.B 29 2007-09-28_B2165-20.gpr U.48.C I.48.C 30 2007-09-28_B2165-23.gpr I.8.B F.8.B 31 2007-09-28_B2165-26.gpr U.24.C I.24.C My code to generate the design matrix for single channel analysis: # seperate channels targets.sc=targetsA2C(targets) u=uniquetargets.sc$Target) f=factortargets.sc$Target, levels=u) design=model.matrix(~0+f) colnames(design)=u The warnings produced by corfit=intraspotCorrelation(MA, design) ... Warning message: NaNs produced in: log(x) Warning message: NaNs produced in: log(x) Warning message: NaNs produced in: log(x) Warning message: NaNs produced in: log(x) > corfit$consensus [1] NaN The warning produced if the design matrix is reduced to one single factor: > design=matrix(rep(1,62), ncol=1) > corfit=intraspotCorrelation(MA, design) Warning message: reml: Max iterations exceeded in: remlscore(y, X, Z) ------------------------------------- Dr Pie M?ller Vector Group Liverpool School of Tropical Medicine Pembroke Place Liverpool L3 5QA UK Tel +44(0) 151 705 3225 Fax +44(0) 151 705 3369 http://www.liv.ac.uk/lstm http://www.ivcc.com
limma limma • 1.1k views
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@gordon-smyth
Last seen 26 minutes ago
WEHI, Melbourne, Australia
Your design, as you've set it up, has insufficient replication to do the statistical analysis. A small number of warnings are not a problem, same as for duplicateCorrelation(). Best wishes Gordon On Fri, 29 Aug 2008, Muller, Pie wrote: > Dear Gordon and Bioconductor list members, > > I am using limma to analyse an unlinked two-colour array experiment > consisting of 31 arrays (with 34,992 spots on each array) and 38 unique > targets. I would like to separate the two channels to perform a separate > channel analysis using the function lmscFit() and tried to follow the > example given in the limma user's guide (see below) but found that the > "intraspotCorrelation()" gives me nothing but NaNs. What could have gone > wrong? There are no NaNs in either the M or A values in my MAlist > object. I suspect there might something wrong with my design matrix as I > noticed that by reducing the design matrix to one single factor such as > "design=matrix(rep(1,62), ncol=1)" I receive a consensus correlation > value back. However, there is still another warning message "reml: Max > iterations exceeded in: remlscore(y, X, Z)". Does anyone have > suggestions as to solve this problem? > > In advance, many thanks for your support! > > Pie > > > My target file: > >> targets > Array Cy3 Cy5 > 1 2007-06-07_B2162-09.gpr F.168.C F.72.B > 2 2007-06-07_B2162-13.gpr U.48.A U.24.A > 3 2007-06-07_B2162-16.gpr F.24.E U.24.C > 4 2007-06-07_B2162-20.gpr F.8.C U.8.C > 5 2007-06-07_B2162-22.gpr U.24.A U.8.A > 6 2007-06-15_B2162-02.gpr F.48.B F.24.D > 7 2007-06-15_B2162-04.gpr U.72.A U.48.A > 8 2007-06-15_B2162-05.gpr F.72.B F.48.B > 9 2007-06-15_B2162-07.gpr F.48.C U.24.C > 10 2007-06-15_B2162-10.gpr F.168.C U.8.B > 11 2007-06-15_B2162-11.gpr U.168.A U.72.A > 12 2007-06-15_B2162-19.gpr F.8.A U.168.A > 13 2007-06-15_B2162-23.gpr F.72.C U.72.C > 14 2007-06-15_B2162-25.gpr F.24.D F.8.B > 15 2007-06-15_B2162-26.gpr F.168.D U.168.C > 16 2007-09-26_B2165-05.gpr I.72.A I.48.A > 17 2007-09-26_B2165-08.gpr I.24.A I.8.A > 18 2007-09-26_B2165-12.gpr I.8.A F.168.B > 19 2007-09-26_B2165-15.gpr I.8.A I.168.B > 20 2007-09-26_B2165-18.gpr I.168.A I.72.A > 21 2007-09-26_B2165-22.gpr I.48.A I.24.A > 22 2007-09-28_B2165-02.gpr U.168.C I.168.C > 23 2007-09-28_B2165-03.gpr I.168.C F.168.C > 24 2007-09-28_B2165-04.gpr I.24.B F.24.D > 25 2007-09-28_B2165-11.gpr I.72.B F.72.B > 26 2007-09-28_B2165-14.gpr U.8.C I.8.C > 27 2007-09-28_B2165-17.gpr U.72.C I.72.C > 28 2007-09-28_B2165-19.gpr I.48.B F.48.B > 29 2007-09-28_B2165-20.gpr U.48.C I.48.C > 30 2007-09-28_B2165-23.gpr I.8.B F.8.B > 31 2007-09-28_B2165-26.gpr U.24.C I.24.C > > > My code to generate the design matrix for single channel analysis: > > # seperate channels > targets.sc=targetsA2C(targets) > u=uniquetargets.sc$Target) > f=factortargets.sc$Target, levels=u) > design=model.matrix(~0+f) > colnames(design)=u > > > The warnings produced by corfit=intraspotCorrelation(MA, design) > > ... > Warning message: > NaNs produced in: log(x) > Warning message: > NaNs produced in: log(x) > Warning message: > NaNs produced in: log(x) > Warning message: > NaNs produced in: log(x) > >> corfit$consensus > [1] NaN > > > The warning produced if the design matrix is reduced to one single factor: > >> design=matrix(rep(1,62), ncol=1) >> corfit=intraspotCorrelation(MA, design) > Warning message: > reml: Max iterations exceeded in: remlscore(y, X, Z) > > ------------------------------------- > > Dr Pie M?ller > Vector Group > Liverpool School of Tropical Medicine > Pembroke Place > Liverpool > L3 5QA > UK > > Tel +44(0) 151 705 3225 > Fax +44(0) 151 705 3369 > > http://www.liv.ac.uk/lstm > http://www.ivcc.com >
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