[BioC] spot weight in limma

Ren Na na at uthscsa.edu
Fri Oct 22 19:44:06 CEST 2004


Hi,
Our 16k mouse array has five types of spot, one of them is "empty" type, 
which means it is just buffer, no oligo in this type of  spots. After I 
used the function read.maimages like:
RG <- read.maimages(targets$FileName, source="spot", wt.fun=wtarea(100))
I checked the weights assigned to the "empty" type of spots, most are 
0.2 to 0.35, but some of them are around 0.5 to 0.7, and  I also can 
find 0.8 or 0.9. Then I  went back to check the image(tiff file). For 
the "empty" type of  spots which have been asigned high weight, I can 
see nothing or just very tiny dot comparing with "oligo" type of spots.
My question is: wtarea downweights spots which are smaller. Those 
"empty" type spots should get weights close to 0. Where I did 
incorrectly ?  I greatly appreciate any advice.
My another question is: we have a batch of slides, it is supposed to 
have 16 k spots. But 600 spots were not printed on those slides at all 
because of machine problem. Here my question is: can I assign weight 0 
to these 600 spots after I get RGList? or I treat them as "empty" spots 
and use the function like above?

Thanks in advance,

Ren



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