[BioC] Normalize background on marray Agilent object
Gordon K Smyth
smyth at wehi.EDU.AU
Thu Jun 20 13:11:44 CEST 2013
Dera Guillermo,
The usual process is to (1) background correct the foreground intensities
with respect to the background, then (2) normalize the M-values
(log-ratios).
For an Agilent two colour array, I do this by:
library(limma)
RG <- read.maimages(files, source="agilent")
RGb <- backgroundCorrect(RG, method="normexp")
MA <- normalizeWithinArrays(RGb, method="loess")
although it is sometimes a good idea to remove positive control probes
before the normalization step.
A recent example using this pipeline is:
http://www.biomedcentral.com/1471-2105/14/165
Best wishes
Gordon
> Date: Wed, 19 Jun 2013 22:38:34 +0200
> From: Guillermo Marco Puche <guillermo.marco at sistemasgenomicos.com>
> To: "bioconductor at r-project.org" <bioconductor at r-project.org>
> Subject: [BioC] Normalize background on marray Agilent object
>
> Hello,
>
> I'm currently trying to normalize rBG values for a marray object.
> Data origin is Agilent dual channel array. I've loaded information with
> readAgilent() function.
>
> What's the correct way to normalize the data? I would like to normalize
> background information first maNorm function manual isn't very
> clarifying for me.
>
> Thanks !
>
> Best regards,
> Guillermo.
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