Manual creation of Affybatch object to study highthrouput qPCR experiments.
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Guest User ★ 13k
@guest-user-4897
Last seen 10.2 years ago
Hi every one. I am using 384 well qPCR array for miRNA study. I wanted to normalize my results using affy. How can I create an affybatch wich contains my Cq values? Thanks a lot. PS. I am new to R/bioconductor. -- output of sessionInfo(): No R command! -- Sent via the guest posting facility at bioconductor.org.
miRNA qPCR affy miRNA qPCR affy • 1.2k views
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@james-w-macdonald-5106
Last seen 2 days ago
United States
Hi Ali, On 10/20/2011 2:49 AM, Ali Mo [guest] wrote: > Hi every one. > > I am using 384 well qPCR array for miRNA study. I wanted to normalize my results using affy. How can I create an affybatch wich contains my Cq values? Thanks a lot. You don't want to do this. Trying to stuff qPCR data into an AffyBatch is asking for trouble. Quantile normalizing qPCR data is asking for trouble as well, IMO, but they are your data. If you want to quantile normalize these data, use normalizeQuantiles() from the limma package. Best. Jim > > PS. I am new to R/bioconductor. > > -- output of sessionInfo(): > > No R command! > > -- > Sent via the guest posting facility at bioconductor.org. > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor -- James W. MacDonald, M.S. Biostatistician Douglas Lab University of Michigan Department of Human Genetics 5912 Buhl 1241 E. Catherine St. Ann Arbor MI 48109-5618 734-615-7826 ********************************************************** 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 Ali, > Hi Ali, > > On 10/20/2011 2:49 AM, Ali Mo [guest] wrote: >> Hi every one. >> >> I am using 384 well qPCR array for miRNA study. I wanted to normalize my >> results using affy. How can I create an affybatch wich contains my Cq >> values? Thanks a lot. > > You don't want to do this. Trying to stuff qPCR data into an AffyBatch > is asking for trouble. Quantile normalizing qPCR data is asking for > trouble as well, IMO, but they are your data. > In case you want to try some other normalisation methods, including e.g. geometric mean which has been recommended for miRNA data, then you might want to have a look at the HTqPCR package. If you type library(HTqPCR) vignette("HTqPCR") that should give you enough instructions on how to start, if you're not too familiar with Bioconductor. There's also a brief comparison of 4-5 different normalisaiton methods, including quantile. HTH \Heidi > If you want to quantile normalize these data, use normalizeQuantiles() > from the limma package. > > Best. > > Jim > > >> >> PS. I am new to R/bioconductor. >> >> -- output of sessionInfo(): >> >> No R command! >> >> -- >> Sent via the guest posting facility at bioconductor.org. >> >> _______________________________________________ >> Bioconductor mailing list >> Bioconductor at r-project.org >> https://stat.ethz.ch/mailman/listinfo/bioconductor >> Search the archives: >> http://news.gmane.org/gmane.science.biology.informatics.conductor > > -- > James W. MacDonald, M.S. > Biostatistician > Douglas Lab > University of Michigan > Department of Human Genetics > 5912 Buhl > 1241 E. Catherine St. > Ann Arbor MI 48109-5618 > 734-615-7826 > > ********************************************************** > Electronic Mail is not secure, may not be read every day, and should not > be used for urgent or sensitive issues > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: > http://news.gmane.org/gmane.science.biology.informatics.conductor >
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james perkins ▴ 300
@james-perkins-2675
Last seen 10.2 years ago
One could use read.qpcr in the new ReadqPCR package, currently in R-devel, to read the data into an eset extending qPCRBatch. You could then normalise with an endogenous control, although I'm not sure how appropriate this is when using miRNA, if you haven't a priori selected a reference. Potentially useful: www.ncbi.nlm.nih.gov/pubmed/18375788 Cheers, Jim On 20 October 2011 07:49, Ali Mo [guest] <guest at="" bioconductor.org=""> wrote: > > Hi every one. > > I am using 384 well qPCR array for miRNA study. I wanted to normalize my results using affy. How can I create an affybatch wich contains my Cq values? Thanks a lot. > > PS. I am new to R/bioconductor. > > ?-- output of sessionInfo(): > > No R command! > > -- > Sent via the guest posting facility at bioconductor.org. > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor >
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