[BioC] How to do normalization without background subtraction
in limmaGUI or limma?
James Wettenhall
wettenhall at wehi.edu.au
Mon Oct 20 07:22:02 MEST 2003
> >I just wonder is there any option to do normalization in limmaGUI or limma
> >WITHOUT background subtraction?
>
> RG <- backgroundCorrect(RG,method="none")
> MA <- normalizeWithinArrays(RG)
>
> Note: you need a more recent version of limma than the BioC release 1.2.
>
> Gordon
Avoiding background subtraction in this way is not yet possible
in limmaGUI via widgets, but you can always edit copies of your
image analysis files (Spot or GenePix or ...) to include columns
of zeroes for the background.
Soon I will be adding some documentation to limmaGUI, to make it
easier to fiddle with the internal limma objects via the
"Evaluate R Code" window.
You can do
RG <- backgroundCorrect(RG,method="none")
But then any existing MA objects will be out of date, so it's
best to start a new analysis, then run the backgroundCorrect
command above, then check the value of MA.Available via the
Evaluate R Code window will show you that the only MA object
which has been automatically created is MAraw
(unnormalized), (not MAwithinArrays, MAbetweenArrays or MAboth),
so you should overwrite this with
MAraw <- MA.RG(RG)
after using background correct "none" to overwrite the default
RG.
Unless you've been doing some serious "hacking" :) the values
in the MA.Available list object seen in the "Evaluate R
Code" window should be consistent with those in the drill-down
status widget.
Regards,
James
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