Problems annotating a gene list
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@quentin-anstee-1257
Last seen 9.6 years ago
Dear List, I am having some problems writing the contents of an MArrayLM object to file and would be grateful for any help. I am processing a 12 chip affy experiment in Bioconductor as follows: Import .CEL files > GCRMA > Filter (kOverA & IQR) to produce an eset file with 4000 probes called 'mySubSet' > Fit linear model using the following script. Libraries loaded: library(simpleaffy) library(genefilter) library(geneplotter) library(multtest) library(affy) library(limma) library(gcrma) library(xtable) library(mouse4302) library(mouse4302cdf) library(annotate) library(annaffy) Script: design <- model.matrix(~ -1+factor(c(1,1,1,1,2,2,2,3,3,3,3,3))) colnames(design) <- c("A", "CONTROL", "B") fit <- lmFit(mySubSet, design) contrastNames<-c("A_CONTROL","B_CONTROL", "A_B") contrast.matrix <- makeContrasts(A-CONTROL, B-CONTROL, A-B, levels=design) fit2 <- contrasts.fit(fit, contrast.matrix) fit2 <- eBayes(fit2) write.fit(fit2,results=NULL,"Annotated_fit2_Table.txt",adjust="none") When I output this table I have all the statistical data as expected but not the gene names and annotation,etc that the help file says should also be included. I guess I must have missed out a step but I'm not sure where I have gone wrong. Any advice much appreciated. Best wishes, Quentin
Annotation gcrma Annotation gcrma • 1.3k views
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@sean-davis-490
Last seen 3 months ago
United States
On 2/24/06 12:32 PM, "Quentin Anstee" <q.anstee at="" imperial.ac.uk=""> wrote: > Dear List, > > I am having some problems writing the contents of an MArrayLM object to file > and would be grateful for any help. I am processing a 12 chip affy > experiment in Bioconductor as follows: > > Import .CEL files > GCRMA > Filter (kOverA & IQR) to produce an eset file > with 4000 probes called 'mySubSet' > Fit linear model using the following > script. > > Libraries loaded: > > library(simpleaffy) > library(genefilter) > library(geneplotter) > library(multtest) > library(affy) > library(limma) > library(gcrma) > library(xtable) > library(mouse4302) > library(mouse4302cdf) > library(annotate) > library(annaffy) > > Script: > > design <- model.matrix(~ -1+factor(c(1,1,1,1,2,2,2,3,3,3,3,3))) > colnames(design) <- c("A", "CONTROL", "B") > fit <- lmFit(mySubSet, design) > contrastNames<-c("A_CONTROL","B_CONTROL", "A_B") > contrast.matrix <- makeContrasts(A-CONTROL, B-CONTROL, A-B, levels=design) > fit2 <- contrasts.fit(fit, contrast.matrix) > fit2 <- eBayes(fit2) > write.fit(fit2,results=NULL,"Annotated_fit2_Table.txt",adjust="none") > > When I output this table I have all the statistical data as expected but not > the gene names and annotation,etc that the help file says should also be > included. I think the help file does not include an "if available" for the gene names and annotation, but it probably should, unless I am mistaken. I think you will have to make your own annotation and attach it to the MArrayLM fit object or use the limma data structures and attach it before running lmFit. Sean
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Hi Sean, Thanks for coming back to me on this one. I have had a look at 'fit2$genes', all it contains are the affy probe IDs as you predicted. Could you describe how to make the annotations and attach them to the MArrayLM file or let me know where this process is described? I have had a look in the limma help files and vignette but have not been able to find a walk through of this procedure. Very many thanks, Quentin > -----Original Message----- > From: Sean Davis [mailto:sdavis2 at mail.nih.gov] > Sent: 24 February 2006 17:42 > To: Quentin Anstee; Bioconductor > Subject: Re: [BioC] Problems annotating a gene list > > > > > On 2/24/06 12:32 PM, "Quentin Anstee" <q.anstee at="" imperial.ac.uk=""> wrote: > > > Dear List, > > > > I am having some problems writing the contents of an > MArrayLM object > > to file and would be grateful for any help. I am processing > a 12 chip > > affy experiment in Bioconductor as follows: > > > > Import .CEL files > GCRMA > Filter (kOverA & IQR) to > produce an eset > > file with 4000 probes called 'mySubSet' > Fit linear model > using the > > following script. > > > > Libraries loaded: > > > > library(simpleaffy) > > library(genefilter) > > library(geneplotter) > > library(multtest) > > library(affy) > > library(limma) > > library(gcrma) > > library(xtable) > > library(mouse4302) > > library(mouse4302cdf) > > library(annotate) > > library(annaffy) > > > > Script: > > > > design <- model.matrix(~ -1+factor(c(1,1,1,1,2,2,2,3,3,3,3,3))) > > colnames(design) <- c("A", "CONTROL", "B") fit <- lmFit(mySubSet, > > design) contrastNames<-c("A_CONTROL","B_CONTROL", "A_B") > > contrast.matrix <- makeContrasts(A-CONTROL, B-CONTROL, A-B, > > levels=design) > > fit2 <- contrasts.fit(fit, contrast.matrix) > > fit2 <- eBayes(fit2) > > > write.fit(fit2,results=NULL,"Annotated_fit2_Table.txt",adjust="none") > > > > When I output this table I have all the statistical data as > expected > > but not the gene names and annotation,etc that the help file says > > should also be included. > > I think the help file does not include an "if available" for > the gene names and annotation, but it probably should, unless > I am mistaken. I think you will have to make your own > annotation and attach it to the MArrayLM fit object or use > the limma data structures and attach it before running lmFit. > > Sean > >
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On 2/24/06 12:58 PM, "Quentin Anstee" <q.anstee at="" imperial.ac.uk=""> wrote: > Hi Sean, > > Thanks for coming back to me on this one. > > I have had a look at 'fit2$genes', all it contains are the affy probe IDs as > you predicted. Could you describe how to make the annotations and attach > them to the MArrayLM file or let me know where this process is described? I > have had a look in the limma help files and vignette but have not been able > to find a walk through of this procedure. It will suffice to make a data.frame in the same order and containing the same number of genes at fit2$genes. If your new data.frame is called gene.data, you just say fit2$genes <- gene.data. You might want to look at the annaffy package and vignette for some ideas about how to ge the information you want from the annotation package. Sean
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Hi Quentin, You might look at limma2annaffy() in the affycoretools package. It will take your design matrix, contrast matrix, exprSet, and MArrayLM objects and output HTML and/or text tables with all the probesets annotated. This package is in the devel repository, so you will not be able to get it using e.g., biocLite() unless you are running R-2.3.0dev. However, it doesn't require the devel version of R, so you can download by hand and install using either the dropdown menu (if you are using win32) or R CMD INSTALL if you are on a Unix variant. HTH, Jim Sean Davis wrote: > > > On 2/24/06 12:58 PM, "Quentin Anstee" <q.anstee at="" imperial.ac.uk=""> wrote: > > >>Hi Sean, >> >>Thanks for coming back to me on this one. >> >>I have had a look at 'fit2$genes', all it contains are the affy probe IDs as >>you predicted. Could you describe how to make the annotations and attach >>them to the MArrayLM file or let me know where this process is described? I >>have had a look in the limma help files and vignette but have not been able >>to find a walk through of this procedure. > > > It will suffice to make a data.frame in the same order and containing the > same number of genes at fit2$genes. If your new data.frame is called > gene.data, you just say fit2$genes <- gene.data. > > You might want to look at the annaffy package and vignette for some ideas > about how to ge the information you want from the annotation package. > > Sean > > _______________________________________________ > Bioconductor mailing list > Bioconductor at stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor -- James W. MacDonald University of Michigan Affymetrix and cDNA Microarray Core 1500 E Medical Center Drive Ann Arbor MI 48109 734-647-5623 ********************************************************** Electronic Mail is not secure, may not be read every day, and should not be used for urgent or sensitive issues.
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Hi Jim, Thanks. That does exactly what I was trying to achieve. The affycoretools package promises to be a great addition to the BioC armoury. All the best, Quentin > -----Original Message----- > From: James W. MacDonald [mailto:jmacdon at med.umich.edu] > Sent: 24 February 2006 22:20 > Cc: Quentin Anstee; Bioconductor > Subject: Re: [BioC] Problems annotating a gene list > > Hi Quentin, > > You might look at limma2annaffy() in the affycoretools > package. It will take your design matrix, contrast matrix, > exprSet, and MArrayLM objects and output HTML and/or text > tables with all the probesets annotated. > > This package is in the devel repository, so you will not be > able to get it using e.g., biocLite() unless you are running > R-2.3.0dev. However, it doesn't require the devel version of > R, so you can download by hand and install using either the > dropdown menu (if you are using win32) or R CMD INSTALL if > you are on a Unix variant. > > HTH, > > Jim > > > Sean Davis wrote: > > > > > > On 2/24/06 12:58 PM, "Quentin Anstee" > <q.anstee at="" imperial.ac.uk=""> wrote: > > > > > >>Hi Sean, > >> > >>Thanks for coming back to me on this one. > >> > >>I have had a look at 'fit2$genes', all it contains are the > affy probe > >>IDs as you predicted. Could you describe how to make the > annotations > >>and attach them to the MArrayLM file or let me know where > this process > >>is described? I have had a look in the limma help files and > vignette > >>but have not been able to find a walk through of this procedure. > > > > > > It will suffice to make a data.frame in the same order and > containing > > the same number of genes at fit2$genes. If your new data.frame is > > called gene.data, you just say fit2$genes <- gene.data. > > > > You might want to look at the annaffy package and vignette for some > > ideas about how to ge the information you want from the > annotation package. > > > > Sean > > > > _______________________________________________ > > Bioconductor mailing list > > Bioconductor at stat.math.ethz.ch > > https://stat.ethz.ch/mailman/listinfo/bioconductor > > > -- > James W. MacDonald > University of Michigan > Affymetrix and cDNA Microarray Core > 1500 E Medical Center Drive > Ann Arbor MI 48109 > 734-647-5623 > > > > ********************************************************** > Electronic Mail is not secure, may not be read every day, and > should not be used for urgent or sensitive issues. >
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