Contrasts matrix for group-means
Since you're using the group-means parametrization, there is now an extra step in the limma pipeline. The coefficients now represent the mean expression level for each of the two groups of samples, so you need to specify a custom contrast to test for differences between the progressive and stable leukemias.
This exercise is part of the course
Differential Expression Analysis with limma in R
Exercise instructions
The ExpressionSet object eset
with the leukemia data and the design matrix you just created (design
) have been loaded in your workspace.
Use
makeContrasts
to define the contraststatus
, which is the difference in the mean expression level between the progressive and stable cancers.Pass the design matrix
design
to the argumentlevels
. This allows the column names ofdesign
(which correspond to the coefficients) to be used in the definitions of the contrasts.
Hands-on interactive exercise
Have a go at this exercise by completing this sample code.
# Load package
library(limma)
# Create a contrasts matrix
cm <- ___(status = ___ - ___,
levels = ___)
# View the contrasts matrix
cm