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Maria Raeder
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10
@maria-raeder-4550
Last seen 10.4 years ago
Dear Mailing List,
I have been struggling for some time with some agilent single channel
arrays, which I believe has been scanned with a earlier version AFE,
because they do not contain the columns Sequence and chr coord, but I
have tried to use the Agi4x44PreProcess package, with some
adjustments, please see below. My main problem now is that I cannot
remove the agilent probe names which are embedded within the
genesymbol column for some genes The reason for doing this is to
prepare files for GSEA analysis. The function for doing this in the
Agi4x44PreProcess package: gsea.files, does not work, porbably due
the the columns I am lacking, and the filter.probes also returns an
error message, probably due to the same reason.
I would be very grateful for any comments and help
Thanks,
Maria
Here is the code :
library("Agi4x44PreProcess")
library("hgug4112a.db")
library("vsn")
library("convert")
library("GO.db")
setwd("/mydirectory")
#reading targets file
targets=read.targets(infile="targets_ec3.txt")
targets[1:10,1:5]
names(targets)
#Many( has skipped them, but included FIleName, Treatment and GErep)
#read in files with LIMMA:
dd <- read.maimages(targets$FileName, source="agilent", columns =
list(G = "gMedianSignal", Gb = "gBGUsed", R = "gProcessedSignal",
Rb = "gBGMedianSignal"), annotation = c("Row", "Col","FeatureNum",
"ControlType","ProbeName","ProbeUID", "GeneName",
"SystematicName", "Description", "gIsWellAboveBG", "gIsFound",
"gIsSaturated", "gIsFeatPopnOL", "gIsFeatNonUnifOL"))
#reads inn 146 arrays)
##########Quality control (skipped)
###########Background correction and normailzation and log 2
transformation:
library(vsn)
ddNORM = BGandNorm(dd, BGmethod = "half", NORMmethod =
"quantile",foreground = "MeanSignal", background = "BGMedianSignal",
offset = 50, makePLOTpre = FALSE, makePLOTpost = FALSE)
#filtering:
ddFILT=filter.probes(ddNORM,
control=TRUE,
wellaboveBG=TRUE,
isfound=TRUE,
wellaboveNEG=TRUE,
sat=TRUE,
PopnOL=TRUE,
NonUnifOL=TRUE,
nas=TRUE,
limWellAbove=75,
limISF=75,
limNEG=75,
limSAT=75,
limPopnOL=75,
limNonUnifOL=75,
limNAS=100,
makePLOT=TRUE,annotation.package="hgug4112a.db",flag.c
ounts=FALSE,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: 43376, 0
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