[BioC] Non-conformable arguments fitting limma contrasts

paul.boutros at utoronto.ca paul.boutros at utoronto.ca
Tue Jul 27 03:43:10 CEST 2004


Hello,

I've come across an error when fitting contrasts with Affy data using limma.  
I'm on:
WinXP SP1
R1.9.1
limma 1.7.2
gcrma 1.1.0
arrays: RAE230A

The error message I'm getting is:
> fit2 <- contrasts.fit(fit1, contrast.matrix);
Error in o %*% RUC^2 : non-conformable arguments
> traceback();
1: contrasts.fit(fit1, contrast.matrix)

Below I've appended  the sequence of commands I'm using, followed by my 
pData(eset), my design and contrast matrices.  

Briefly, what I have are the following factors:
Strain: four levels (HW, LE, LnC, LnA)
Time: two levels (hour3, hour19)
Treatment: two levels (TCDD, COil)
AHR Genotype: two levels (HW, WT)

Where I believe I may have gone wrong is explicitly including the AHR genotype 
in the model.  This factor can be determined by the strain: two strains (HW, 
LnA) always have AHR = HW while the other two strains always have AHR = WT.  
Including both surely feels like double-fitting to me.  On the other hand, the 
AHR genotype is *not* the only factor differing between the strains which was my 
motivation for including it.

As always, I'm open to any comments/suggestions/corrections.
Paul


### Start Command Sequence ###
eset <- ReadAffy(filenames=cel.files, phenoData="phenodata.txt");
eset <- gcrma(eset);

# create a design matrix:
design <- model.matrix(~-1 + Strain + Treatment + Time + AHR + AHR:Treatment + 
AHR:Time, pData(eset));
colnames(design) <- c("HW", "LE", "LnA", "LnC", "TCDD", "Time", "WT", "WT.TCDD", 
"WT.Time");

# make a contrast matrix
contrast.matrix <- makeContrasts(TCDD, Time, -WT, -WT.TCDD, -WT.Time, 
levels=design);

# fit the model
fit1 <- lmFit(eset, design);
fit2 <- contrasts.fit(fit1, contrast.matrix);
fit3 <- eBayes(fit2);
### End Command Sequence ###

> pData(eset);
                            Strain Treatment   Time AHR
RAE230A_021304_IM01T_LH.CEL     HW      TCDD  Hour3  HW
RAE230A_021304_IM02T_LH.CEL     HW      TCDD  Hour3  HW
RAE230A_021304_IM03T_LH.CEL     HW      TCDD  Hour3  HW
RAE230A_021304_IM04T_LH.CEL     HW      TCDD  Hour3  HW
RAE230A_021304_IM05T_LH.CEL     LE      TCDD  Hour3  WT
RAE230A_021304_IM06T_LH.CEL     LE      TCDD  Hour3  WT
RAE230A_021304_IM07T_LH.CEL     LE      TCDD  Hour3  WT
RAE230A_021304_IM08T_LH.CEL     LE      TCDD  Hour3  WT
RAE230A_043003_IM01T_LH.CEL     LE      TCDD Hour19  WT
RAE230A_043003_IM02T_LH.CEL     LE      COil Hour19  WT
RAE230A_043003_IM03T_LH.CEL     HW      COil Hour19  HW
RAE230A_043003_IM04T_LH.CEL     HW      TCDD Hour19  HW
RAE230A_060403_IM01T_LH.CEL     LE      COil Hour19  WT
RAE230A_060403_IM02T_LH.CEL     LE      COil Hour19  WT
RAE230A_060403_IM03T_LH.CEL     LE      COil Hour19  WT
RAE230A_060403_IM04T_LH.CEL     HW      COil Hour19  HW
RAE230A_060403_IM05T_LH.CEL     HW      COil Hour19  HW
RAE230A_060403_IM06T_LH.CEL     LE      TCDD Hour19  WT
RAE230A_060403_IM07T_LH.CEL     LE      TCDD Hour19  WT
RAE230A_060403_IM08T_LH.CEL     LE      TCDD Hour19  WT
RAE230A_060403_IM09T_LH.CEL     HW      TCDD Hour19  HW
RAE230A_060403_IM10T_LH.CEL     HW      TCDD Hour19  HW
RAE230A_070303_IM01T_LH.CEL     HW      TCDD Hour19  HW
RAE230A_070303_IM02T_LH.CEL     HW      COil Hour19  HW
RAE230A_102803_IM01T_LH.CEL    LnA      COil Hour19  HW
RAE230A_102803_IM02T_LH.CEL    LnA      COil Hour19  HW
RAE230A_102803_IM03T_LH.CEL    LnA      COil Hour19  HW
RAE230A_102803_IM04T_LH.CEL    LnA      COil Hour19  HW
RAE230A_102803_IM05T_LH.CEL    LnA      TCDD Hour19  HW
RAE230A_102803_IM06T_LH.CEL    LnA      TCDD Hour19  HW
RAE230A_102803_IM07T_LH.CEL    LnA      TCDD Hour19  HW
RAE230A_102803_IM08T_LH.CEL    LnC      COil Hour19  WT
RAE230A_102803_IM09T_LH.CEL    LnC      COil Hour19  WT
RAE230A_102803_IM10T_LH.CEL    LnC      COil Hour19  WT
RAE230A_102803_IM11T_LH.CEL    LnC      COil Hour19  WT
RAE230A_102803_IM12T_LH.CEL    LnC      TCDD Hour19  WT
RAE230A_102803_IM13T_LH.CEL    LnC      TCDD Hour19  WT
RAE230A_102803_IM14T_LH.CEL    LnC      TCDD Hour19  WT
RAE230A_102803_IM15T_LH.CEL    LnC      TCDD Hour19  WT
RAE230A_102803_IM16T_LH.CEL    LnA      TCDD Hour19  HW

> design;

                            HW LE LnA LnC TCDD Time WT WT.TCDD WT.Time
RAE230A_021304_IM01T_LH.CEL  1  0   0   0    1    1  0       0       0
RAE230A_021304_IM02T_LH.CEL  1  0   0   0    1    1  0       0       0
RAE230A_021304_IM03T_LH.CEL  1  0   0   0    1    1  0       0       0
RAE230A_021304_IM04T_LH.CEL  1  0   0   0    1    1  0       0       0
RAE230A_021304_IM05T_LH.CEL  0  1   0   0    1    1  1       1       1
RAE230A_021304_IM06T_LH.CEL  0  1   0   0    1    1  1       1       1
RAE230A_021304_IM07T_LH.CEL  0  1   0   0    1    1  1       1       1
RAE230A_021304_IM08T_LH.CEL  0  1   0   0    1    1  1       1       1
RAE230A_043003_IM01T_LH.CEL  0  1   0   0    1    0  1       1       0
RAE230A_043003_IM02T_LH.CEL  0  1   0   0    0    0  1       0       0
RAE230A_043003_IM03T_LH.CEL  1  0   0   0    0    0  0       0       0
RAE230A_043003_IM04T_LH.CEL  1  0   0   0    1    0  0       0       0
RAE230A_060403_IM01T_LH.CEL  0  1   0   0    0    0  1       0       0
RAE230A_060403_IM02T_LH.CEL  0  1   0   0    0    0  1       0       0
RAE230A_060403_IM03T_LH.CEL  0  1   0   0    0    0  1       0       0
RAE230A_060403_IM04T_LH.CEL  1  0   0   0    0    0  0       0       0
RAE230A_060403_IM05T_LH.CEL  1  0   0   0    0    0  0       0       0
RAE230A_060403_IM06T_LH.CEL  0  1   0   0    1    0  1       1       0
RAE230A_060403_IM07T_LH.CEL  0  1   0   0    1    0  1       1       0
RAE230A_060403_IM08T_LH.CEL  0  1   0   0    1    0  1       1       0
RAE230A_060403_IM09T_LH.CEL  1  0   0   0    1    0  0       0       0
RAE230A_060403_IM10T_LH.CEL  1  0   0   0    1    0  0       0       0
RAE230A_070303_IM01T_LH.CEL  1  0   0   0    1    0  0       0       0
RAE230A_070303_IM02T_LH.CEL  1  0   0   0    0    0  0       0       0
RAE230A_102803_IM01T_LH.CEL  0  0   1   0    0    0  0       0       0
RAE230A_102803_IM02T_LH.CEL  0  0   1   0    0    0  0       0       0
RAE230A_102803_IM03T_LH.CEL  0  0   1   0    0    0  0       0       0
RAE230A_102803_IM04T_LH.CEL  0  0   1   0    0    0  0       0       0
RAE230A_102803_IM05T_LH.CEL  0  0   1   0    1    0  0       0       0
RAE230A_102803_IM06T_LH.CEL  0  0   1   0    1    0  0       0       0
RAE230A_102803_IM07T_LH.CEL  0  0   1   0    1    0  0       0       0
RAE230A_102803_IM08T_LH.CEL  0  0   0   1    0    0  1       0       0
RAE230A_102803_IM09T_LH.CEL  0  0   0   1    0    0  1       0       0
RAE230A_102803_IM10T_LH.CEL  0  0   0   1    0    0  1       0       0
RAE230A_102803_IM11T_LH.CEL  0  0   0   1    0    0  1       0       0
RAE230A_102803_IM12T_LH.CEL  0  0   0   1    1    0  1       1       0
RAE230A_102803_IM13T_LH.CEL  0  0   0   1    1    0  1       1       0
RAE230A_102803_IM14T_LH.CEL  0  0   0   1    1    0  1       1       0
RAE230A_102803_IM15T_LH.CEL  0  0   0   1    1    0  1       1       0
RAE230A_102803_IM16T_LH.CEL  0  0   1   0    1    0  0       0       0
attr(,"assign")
[1] 1 1 1 1 2 3 4 5 6
attr(,"contrasts")
attr(,"contrasts")$Strain
[1] "contr.treatment"

attr(,"contrasts")$Treatment
[1] "contr.treatment"

attr(,"contrasts")$Time
[1] "contr.treatment"

attr(,"contrasts")$AHR
[1] "contr.treatment"

> contrast.matrix;
        TCDD Time -WT -WT.TCDD -WT.Time
HW         0    0   0        0        0
LE         0    0   0        0        0
LnA        0    0   0        0        0
LnC        0    0   0        0        0
TCDD       1    0   0        0        0
Time       0    1   0        0        0
WT         0    0  -1        0        0
WT.TCDD    0    0   0       -1        0
WT.Time    0    0   0        0       -1



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