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wefelmeyer@web.de
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@wefelmeyerwebde-1347
Last seen 10.2 years ago
Dear list,
I have got 2x3 arrays (3 wildtype, 3 mutant) of the type MOE 430 2.0
(mouse genome).
Since there are so many different approaches to normalization, I do
not really know which one to choose.
I thought about using GCRMA, since it is said to be good for low
expressed genes and I have quite a lot of those (double knockout
experiment).
Also, I thought GCRMA would be better than, say, VSN for my design,
since parameter estimation out of 3 chips doesn't seem to be very
trustworthy.
I got a few questions concerning GCRMA, though. When looking at the
histogram of the variances (across all arrays), I see a clear bimodal
distribution. After searching in the mail archives here, I found some
hints at GCRMA being only optimised for a certain human chip. Is that
true? Should I rather be using another method? Any suggestions on
this?
In general, the quality control plots of the gcrma normalized values
seem okay to me. I looked at variance to mean expression value,
wildtype versus mutant variances scatterplot, boxplot and others.
When comparing the histograms of the expression values or variances to
other normalization methods (rma, vsn, mas5), I get obviously
different distributions. Is that good or bad? Any suggestions on how
to decide on this?
Thanks a lot for helping out.
Best, ww
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