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Tiandao Li
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260
@tiandao-li-2372
Last seen 10.3 years ago
Hello,
I am using limma for 2-color microarray data analysis. I have some
questions regarding MA, design, and contrasts.
1. I have one MA file including all my experiments. Rows of MA
correspond
to spots and columns to individual experimental file. Columns were
listed
as the increasing order of file names, since I used barcodes as file
names. Then after reading in the target file and creating design
matrix, I
used the following to caluculate correlation between duplicates.
corfit <- duplicateCorrelation(MA,design,ndups=4) # A slow
computation!
corfit$consensus.correlation
However if columns of MA2 are listed following the order of
target$FileName, corfit2$consensus.correlation value is different from
one
of MA. Which one should I use in the next anlysis?
2. I have one MA file including all my experiments, and also an all-
in-one
contrast matrix including different contrasts (related or un-related).
Should I use this all-in-one contrast matrix for linear model to find
the
differentially expressed genes? This doesn't sound right. Or I use
subset
of MA for and only for related one or more contrasts, and use all-in-
one
MA only necessary. Which one is better?
Thanks in advance,
Tiandao