[BioC] help with Agilent microarrays
Alberto Goldoni
alberto.goldoni1975 at gmail.com
Thu Sep 2 15:32:22 CEST 2010
Hi to everybody,
i'm analysing 2 color agilent microarrays (rgug4130a.db) and my
targets.txt file is:
"Name" "FileName" "Cy3" "Cy5" "GErep"
"prova01" "p01.txt" "ref" "wt" 1
"prova02" "p02.txt" "ref" "wt" 1
"prova03" "p03.txt" "ref" "wt" 1
"prova04" "p04.txt" "ref" "ko" 2
"prova05" "p05.txt" "ref" "ko" 2
"prova06" "p06.txt" "ref" "ko" 2
now i have obtainet the data with "limma" and for the analysis i'm
using "Agi4x44PreProcess" package.
All works fine until the normalization:
RG <- read.maimages(targets, source="agilent")
> RG_NORM = BGandNorm(RG, BGmethod = "half", NORMmethod = "quantile", foreground = "MeanSignal", background = "BGMedianSignal", offset = 50, makePLOTpre = FALSE, makePLOTpost = FALSE)
BACKGROUND CORRECTION AND NORMALIZATION
foreground: MeanSignal
background: BGMedianSignal
BGmethod: half
NORMmethod: quantile
OUTPUT in log-2 scale
------------------------------------------------------
but when i'm tryng to do the filtering i obtain the follow error:
> RGFILT = filter.probes(RG_NORM, control = TRUE, wellaboveBG = TRUE, isfound = TRUE, wellaboveNEG = TRUE, sat = TRUE, PopnOL = TRUE, NonUnifOL = T, nas = TRUE, limWellAbove = 75, limISF = 75, limNEG = 75, limSAT = 75, limPopnOL = 75, limNonUnifOL = 75, limNAS = 100, makePLOT = F, annotation.package = "rgug4130a.db", flag.counts = T, targets)
FILTERING PROBES BY FLAGS
FILTERING BY ControlType FLAG
Error in data.frame(PROBE_ID, as.character(probe.chr),
as.character(probe.seq), :
arguments imply differing number of rows: 20500, 0
someone can help me???
best regards
--
-----------------------------------------------------
Dr. Alberto Goldoni
Parma, Italy
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