Thanks a lot for the answer!
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I would like to find out differentially expressed genes in 4 different groups; however, each group has triplicates. So, what is the correct way to follow in this scenario in order to compare the groups between themselves? I guess i can do it if they were single, but being in triplicate confuses me a bit.
Data is from Illumina HumanHT-12 v4 Expression BeadChip, i exported from GenomeStudio v2011.
Thanks a lot!
Take a look at Chapter 7 (Linear Models Overview) of the limma users guide. It has some good examples. To compare 4 groups consisting of triplicate samples you would do something like the following:
library(limma)
design <- model.matrix(~ 0+factor(c(1,1,1,2,2,2,3,3,3,4,4,4)))
colnames(design) <- c("group1", "group2", "group3", "group4")
# call lmFit on your data
fit <- lmFit(yourdata, design)
# create you comparisons of interest
contrast.matrix <- makeContrasts(group2-group1, group3-group2, group3-group1, group4-group1, group4-group3, group4-group2, levels=design)
fit2 <- contrasts.fit(fit, contrast.matrix)
fit2 <- eBayes(fit2)
# fit 2 now contains all your comparisons
head(fit2$coef)
Thanks a lot for the answer!
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