[BioC] LimmaGUI Spot Quality
James Wettenhall
wettenhall at wehi.edu.au
Fri Jul 2 17:41:52 CEST 2004
Liz,
On Fri, 2 Jul 2004, Elizabeth Brooke-Powell wrote:
> adjust the model appropriately. I am not sure in this case that pretending
> to have GenePix will work as the numbers are not a simple 0 or 1 (good or
No, sorry I didn't mean to imply that you would be able to just
use the GenePix option in limmaGUI as is. I just thought it
might by helpful for you to learn how weights can be defined
(for _GenePix_ data), based on GenePix spot flags. Notice that
the weights we define for the GenePix flags are between 0 and
1, just as your "quality weights" already are. But after we
process GenePix data, the number of _different_ values in the
weights column would be small, e.g. in this weights vector:
(1,1,1,1,0.1,1,1,1,1,1,1,0,1,1,1,1,1,0.1,0.1,1,1,1,1,1),
there are only three _different_ weight values (0, 0.1 and 1),
whereas for your data, the column of weights (between 0 and 1)
could contain lots of different weight values between 0 and 1
for the different genes.
I don't think you have told us the column name of this
quality weight yet.
Maybe you should ask the statistician who designed this
quality weighting how he/she intended that it be used in
normalization. But it can probably be used directly in
limma's normalization, and all you would have to do is tell us
the appropriate column names which limma would need to read in
for your data (Rf, Rb, Gf, Gb and Spot-Quality-Weighting) and
then we can add an option to limma/limmaGUI to allow it to read
in the appropriate columns for BlueFuse including the quality
weights.
There are no plans at the moment to add a custom-dialog to
limmaGUI for reading in an arbitrary column of weights from your
raw image-analysis files. But if you want to start combining
the command-line interface with the GUI interface, you could
read the weights into RG$weights in limmaGUIenvironment. Then
they would be automatically used for normalization.
(1) From the R console :
RG <- get("RG",envir=limmaGUIenvironment)
names(RG)
RG$weights <- ...
names(RG)
assign("RG",RG,limmaGUIenvironment)
OR
(2) From the "Evaluate R Code menu:
RG$weights <- ...
(In case (2), when using the "Evaluate R Code" menu, your R
commands are automatically evaluated in limmaGUIenvironment
which contains all of your microarray data objects used by
limmaGUI.)
Regards,
James
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