HTqPCR - limmaCtData using samples with different number of replicates?
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@fletez-brant-christopher-nihvrc-c-5360
Last seen 10.3 years ago
Dear List, I am using HTqPCR to analyze Fluidigm data (specifically, the 96*96 format). I have been using specifically limmaCtData to look for differential expression between groups, always having the same number of replicates. However, I currently have a dataset of several groups with 3 replicates and 1 group with 2 replicates. As far as I can tell, limmaCtData expects the number of replicates to be the same between groups (i.e. there is only one 'ndups' parameter). Has anyone else encountered this issue? Thank you, Kipper Fletez-Brant
HTqPCR HTqPCR • 1.3k views
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Heidi Dvinge ★ 2.0k
@heidi-dvinge-2195
Last seen 10.3 years ago
Hi Kipper, > Dear List, > > I am using HTqPCR to analyze Fluidigm data (specifically, the 96*96 > format). I have been using specifically limmaCtData to look for > differential expression between groups, always having the same number of > replicates. However, I currently have a dataset of several groups with 3 > replicates and 1 group with 2 replicates. As far as I can tell, > limmaCtData expects the number of replicates to be the same between groups > (i.e. there is only one 'ndups' parameter). Has anyone else encountered > this issue? > ndups actually refers to the number of replicate *features*, i.e. genes, on your plate. As long as you use the same platform for all your samples, this should be constant. The number of samples within each group is indicated using the design matrix in limmaCtData, and the parameter 'groups' in ttestCtData. So in limmaCtData you can just use a design matrix along the lines of: > samples <- rep(c("treat1", "treat2", "treat3", "control"), c(3,2,3,3)) > design <- model.matrix(~0+samples) You then indicate your comparisons of interest in the contrast matrix, and use both matrices in your call to limmaCtData. If I've misunderstood you and you actually have a variable number of replicates of each gene, then I'm afraid limmaCtData isn't capable of handling this with ndups. HTH \Heidi > Thank you, > Kipper Fletez-Brant > > _______________________________________________ > 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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Hi Heidi, Thanks for setting me straight - I in fact have a panel of 96 unrepeated features, used to produce experiments with a variable number of replicates per group. This works very well. Thanks! On 6/26/12 12:14 PM, "Heidi Dvinge" <heidi at="" ebi.ac.uk=""> wrote: >Hi Kipper, > >> Dear List, >> >> I am using HTqPCR to analyze Fluidigm data (specifically, the 96*96 >> format). I have been using specifically limmaCtData to look for >> differential expression between groups, always having the same number of >> replicates. However, I currently have a dataset of several groups with >>3 >> replicates and 1 group with 2 replicates. As far as I can tell, >> limmaCtData expects the number of replicates to be the same between >>groups >> (i.e. there is only one 'ndups' parameter). Has anyone else encountered >> this issue? >> >ndups actually refers to the number of replicate *features*, i.e. genes, >on your plate. As long as you use the same platform for all your samples, >this should be constant. > >The number of samples within each group is indicated using the design >matrix in limmaCtData, and the parameter 'groups' in ttestCtData. So in >limmaCtData you can just use a design matrix along the lines of: > >> samples <- rep(c("treat1", "treat2", "treat3", "control"), c(3,2,3,3)) >> design <- model.matrix(~0+samples) > >You then indicate your comparisons of interest in the contrast matrix, and >use both matrices in your call to limmaCtData. > >If I've misunderstood you and you actually have a variable number of >replicates of each gene, then I'm afraid limmaCtData isn't capable of >handling this with ndups. > >HTH >\Heidi > >> Thank you, >> Kipper Fletez-Brant >> >> _______________________________________________ >> 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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