Inconsistency in RMA results from 'affy' and results from 'oligo'
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Peng Yu ▴ 940
@peng-yu-3586
Last seen 9.7 years ago
Hi, I have run the two different R script to do RMA. Neither of them gives me any error messages. However, the RMA results are very different---they have very different number of lines. I don't know which one I should believe. Or neither of them is correct. It might be due to the difference in the cdf file used. Would you please point to me how to figure out the problem? $ Rscript probe2expr_affy.R > library(affy) Loading required package: Biobase Loading required package: methods Welcome to Bioconductor Vignettes contain introductory material. To view, type 'openVignette()'. To cite Bioconductor, see 'citation("Biobase")' and for packages 'citation(pkgname)'. > Data <- ReadAffy() > eset <- rma(Data) Background correcting Normalizing Calculating Expression > write.exprs(eset, file="gene_expr_affy.txt", sep="\t") > $ Rscript probe2expr_oligo.R > library(oligo) Loading required package: oligoClasses Loading required package: Biobase Loading required package: methods Welcome to Bioconductor Vignettes contain introductory material. To view, type 'openVignette()'. To cite Bioconductor, see 'citation("Biobase")' and for packages 'citation(pkgname)'. Loading required package: preprocessCore Welcome to oligo version 1.8.1 > data<-read.celfiles(list.celfiles()) Loading required package: pd.mogene.1.0.st.v1 Loading required package: RSQLite Loading required package: DBI Platform design info loaded. Reading in : koA-mth_HZ_5238_MST1_19389.cel Reading in : koB-mth_HZ_5238_MST1_19390.cel Reading in : koC-mth_HZ_5238_MST1_19391.cel Reading in : koD-mth_HZ_5238_MST1_19392.cel Reading in : wt1-mth_HZ_5238_MST1_19385.cel Reading in : wt2-mth_HZ_5238_MST1_19386.cel Reading in : wt3-mth_HZ_5238_MST1_19387.cel Reading in : wt4-mth_HZ_5238_MST1_19388.cel > eset<-rma(data) Background correcting Normalizing Calculating Expression > write.exprs(eset, file="gene_expr_oligo.txt", sep="\t") > $ wc gene_expr_affy.txt gene_expr_oligo.txt 34761 312848 5002519 gene_expr_affy.txt 234591 2111318 33763075 gene_expr_oligo.txt 269352 2424166 38765594 total BTW, before I run 'Rscript probe2expr_affy.R'. I downloaded MoGene-1_0-st-v1.r3.unsupported-cdf and I run the following script. Then, I install the generated package 'mogene10stv1cdf'. $ cat make_cdf_package.R library(makecdfenv) make.cdf.package("MoGene-1_0-st-v1.r3.cdf", packagename="mogene10stv1cdf", species="Mus_musculus") Regards, Peng
cdf oligo cdf oligo • 1.0k views
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@benilton-carvalho-1375
Last seen 4.1 years ago
Brazil/Campinas/UNICAMP
Dear Peng, I can speak for oligo and the annotation package used by it. The current release of oligo summarizes to the probeset level. The next release of oligo and annotation packages will allow you to summarize to the gene level. In your particular case, the count you'll get is roughly 35K. The updated packages have already been submitted to BioC and soon should show up on the devel branch. Best wishes, b On Aug 10, 2009, at 6:39 PM, Peng Yu wrote: > Hi, > > I have run the two different R script to do RMA. Neither of them gives > me any error messages. However, the RMA results are very > different---they have very different number of lines. I don't know > which one I should believe. Or neither of them is correct. It might be > due to the difference in the cdf file used. Would you please point to > me how to figure out the problem? > > $ Rscript probe2expr_affy.R >> library(affy) > Loading required package: Biobase > Loading required package: methods > > Welcome to Bioconductor > > Vignettes contain introductory material. To view, type > 'openVignette()'. To cite Bioconductor, see > 'citation("Biobase")' and for packages 'citation(pkgname)'. > >> Data <- ReadAffy() >> eset <- rma(Data) > Background correcting > Normalizing > Calculating Expression >> write.exprs(eset, file="gene_expr_affy.txt", sep="\t") >> > > $ Rscript probe2expr_oligo.R >> library(oligo) > Loading required package: oligoClasses > Loading required package: Biobase > Loading required package: methods > > Welcome to Bioconductor > > Vignettes contain introductory material. To view, type > 'openVignette()'. To cite Bioconductor, see > 'citation("Biobase")' and for packages 'citation(pkgname)'. > > Loading required package: preprocessCore > Welcome to oligo version 1.8.1 >> data<-read.celfiles(list.celfiles()) > Loading required package: pd.mogene.1.0.st.v1 > Loading required package: RSQLite > Loading required package: DBI > Platform design info loaded. > Reading in : koA-mth_HZ_5238_MST1_19389.cel > Reading in : koB-mth_HZ_5238_MST1_19390.cel > Reading in : koC-mth_HZ_5238_MST1_19391.cel > Reading in : koD-mth_HZ_5238_MST1_19392.cel > Reading in : wt1-mth_HZ_5238_MST1_19385.cel > Reading in : wt2-mth_HZ_5238_MST1_19386.cel > Reading in : wt3-mth_HZ_5238_MST1_19387.cel > Reading in : wt4-mth_HZ_5238_MST1_19388.cel >> eset<-rma(data) > Background correcting > Normalizing > Calculating Expression >> write.exprs(eset, file="gene_expr_oligo.txt", sep="\t") >> > > $ wc gene_expr_affy.txt gene_expr_oligo.txt > 34761 312848 5002519 gene_expr_affy.txt > 234591 2111318 33763075 gene_expr_oligo.txt > 269352 2424166 38765594 total > > BTW, before I run 'Rscript probe2expr_affy.R'. I downloaded > MoGene-1_0-st-v1.r3.unsupported-cdf and I run the following script. > Then, I install the generated package 'mogene10stv1cdf'. > $ cat make_cdf_package.R > library(makecdfenv) > make.cdf.package("MoGene-1_0-st-v1.r3.cdf", > packagename="mogene10stv1cdf", species="Mus_musculus") > > Regards, > Peng > > _______________________________________________ > Bioconductor mailing list > Bioconductor at stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor
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Hi Benilton, One thing I don't understand isf why 'oligo' didin't require me to install the .cdf file, but 'affy' required me to install the .cdf file. I am curious that how 'oligo' figure how where the correct .cdf file is and use it? Regards, Peng On Mon, Aug 10, 2009 at 5:25 PM, Benilton Carvalho<bcarvalh at="" jhsph.edu=""> wrote: > Dear Peng, > > I can speak for oligo and the annotation package used by it. > > The current release of oligo summarizes to the probeset level. The next > release of oligo and annotation packages will allow you to summarize to the > gene level. In your particular case, the count you'll get is roughly 35K. > > The updated packages have already been submitted to BioC and soon should > show up on the devel branch. > > Best wishes, > > b > > On Aug 10, 2009, at 6:39 PM, Peng Yu wrote: > >> Hi, >> >> I have run the two different R script to do RMA. Neither of them gives >> me any error messages. However, the RMA results are very >> different---they have very different number of lines. I don't know >> which one I should believe. Or neither of them is correct. It might be >> due to the difference in the cdf file used. Would you please point to >> me how to figure out the problem? >> >> $ Rscript probe2expr_affy.R >>> >>> library(affy) >> >> Loading required package: Biobase >> Loading required package: methods >> >> Welcome to Bioconductor >> >> Vignettes contain introductory material. To view, type >> 'openVignette()'. To cite Bioconductor, see >> 'citation("Biobase")' and for packages 'citation(pkgname)'. >> >>> Data <- ReadAffy() >>> eset <- rma(Data) >> >> Background correcting >> Normalizing >> Calculating Expression >>> >>> write.exprs(eset, file="gene_expr_affy.txt", sep="\t") >>> >> >> $ Rscript probe2expr_oligo.R >>> >>> library(oligo) >> >> Loading required package: oligoClasses >> Loading required package: Biobase >> Loading required package: methods >> >> Welcome to Bioconductor >> >> Vignettes contain introductory material. To view, type >> 'openVignette()'. To cite Bioconductor, see >> 'citation("Biobase")' and for packages 'citation(pkgname)'. >> >> Loading required package: preprocessCore >> Welcome to oligo version 1.8.1 >>> >>> data<-read.celfiles(list.celfiles()) >> >> Loading required package: pd.mogene.1.0.st.v1 >> Loading required package: RSQLite >> Loading required package: DBI >> Platform design info loaded. >> Reading in : koA-mth_HZ_5238_MST1_19389.cel >> Reading in : koB-mth_HZ_5238_MST1_19390.cel >> Reading in : koC-mth_HZ_5238_MST1_19391.cel >> Reading in : koD-mth_HZ_5238_MST1_19392.cel >> Reading in : wt1-mth_HZ_5238_MST1_19385.cel >> Reading in : wt2-mth_HZ_5238_MST1_19386.cel >> Reading in : wt3-mth_HZ_5238_MST1_19387.cel >> Reading in : wt4-mth_HZ_5238_MST1_19388.cel >>> >>> eset<-rma(data) >> >> Background correcting >> Normalizing >> Calculating Expression >>> >>> write.exprs(eset, file="gene_expr_oligo.txt", sep="\t") >>> >> >> $ wc gene_expr_affy.txt gene_expr_oligo.txt >> ?34761 ? 312848 ?5002519 gene_expr_affy.txt >> 234591 ?2111318 33763075 gene_expr_oligo.txt >> 269352 ?2424166 38765594 total >> >> BTW, before I run 'Rscript probe2expr_affy.R'. I downloaded >> MoGene-1_0-st-v1.r3.unsupported-cdf and I run the following script. >> Then, I install the generated package 'mogene10stv1cdf'. >> $ cat make_cdf_package.R >> library(makecdfenv) >> make.cdf.package("MoGene-1_0-st-v1.r3.cdf", >> packagename="mogene10stv1cdf", species="Mus_musculus") >> >> Regards, >> Peng >> >> _______________________________________________ >> Bioconductor mailing list >> Bioconductor at stat.math.ethz.ch >> https://stat.ethz.ch/mailman/listinfo/bioconductor >> Search the archives: >> http://news.gmane.org/gmane.science.biology.informatics.conductor > >
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