[BioC] LIMMA: paired samples and common reference?
Enx Enx
enx23 at yahoo.com
Mon Aug 15 08:42:16 CEST 2011
Hello,
I havea an experiment with mice done on Affymetrix and it looks like this:
FileName Treatment Batch MouseID
===============================
file1 control 1 101
file2 control 1 102
file3 control 1 103
file4 treatA 1 104
file5 treatA 1 105
file6 treatA 1 106
file7 control 2 107
file8 control 2 108
file9 control 2 109
file10 treatA 2 110
file11 treatA 2 111
file12 treatA 2 112
file13 control 3 113
file14 control 3 114
file15 control 3 115
file16 treatB 3 116
file17 treatB 3 117
file18 treatB 3 118
file19 control 4 119
file20 control 4 120
file21 control 4 121
file22 treatB 4 122
file23 treatB 4 123
file24 treatB 4 124
The controls from all batches are (should be) biologically the same and I am looking for differentially expressed genes in:
a) treatB versus treatA,
b) treatA versus control,
c) treatB versus control.
Are the design matrix and contrast matrix as follows?
factor_treatment = factor(Treatment, levels=C("control","treatA","treatB"))
factor_batch = factor(Batch, levels=C("1","2","3","4"))
design = model.matrix(~0 + factor_batch + factor_treatment)
fit = lmFit(eset, design)
cont.matrix = makeContrast(
treatBvsA = (treatB - treatA),
treatAvsControl = (treatA - control),
treatBvsControl = (treatB - control),
levels = design)
fit2 = lmFit(fit, cont.matrix)
fit2 = eBayes(fit2)
Cheers,
Daniel Berger
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