limma question
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@ivanborozanutorontoca-704
Last seen 9.6 years ago
hi there, I would like to estimate the effect on gene expression levels of two factors Age and activity (each with 4 levels) using Limma. for my design matrix i have dataB<-data.frame(samples = arrays,Age=factor(FFAge),activity = factor(FFfibrosis)) design<-model.matrix(~activity*Age, data=dataB) (samples contain the names of my arrays) fit <- lm.series(MANormBetween$M,design) toptable(coef=2,num=10,genelist=gal,fit=fit,adjust="fdr",sort.by="P") I would like to know to which effect the quoted P.values in toptable() correspond to ? Also i would like to know how to extract the P.value for the activity:Age effect. all the best.
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@james-wettenhall-153
Last seen 9.6 years ago
Ivan, > toptable(coef=2,num=10,genelist=gal,fit=fit,adjust="fdr",sort.by="P") > > I would like to know to which effect the quoted P.values in toptable() > correspond to ? coef=2 means the effect described by the second column of your design matrix. Hope this helps, James
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@gordon-smyth
Last seen 13 minutes ago
WEHI, Melbourne, Australia
At 01:28 AM 3/04/2004, ivan.borozan@utoronto.ca wrote: >hi there, > >I would like to estimate the effect on gene expression levels of two >factors Age >and activity (each with 4 levels) using Limma. > >for my design matrix i have > >dataB<-data.frame(samples = arrays,Age=factor(FFAge),activity = >factor(FFfibrosis)) > >design<-model.matrix(~activity*Age, data=dataB) > >(samples contain the names of my arrays) > >fit <- lm.series(MANormBetween$M,design) > >toptable(coef=2,num=10,genelist=gal,fit=fit,adjust="fdr",sort.by="P") > >I would like to know to which effect the quoted P.values in toptable() >correspond to ? Look at colnames(design). You've asked for the 2nd coefficient. >Also i would like to know how to extract the P.value for the activity:Age >effect. The activity:effect interaction is on 9 degrees of freedom and I assume that you understand that you need an F-statistic rather than a t-test statistic to test this composite hypothesis. Your fitted factorial model has 16 parameters of which the last 9 are interaction terms. Using limma 1.5.2 or later, you can use fit <- lmFit(MANormBetween@M, design) cont.matrix <- rbind( matrix(0,7,9), diag(9) ) # pick out last 9 coefficients fit <- contrasts.fit(fit, cont.matrix) fit <- eBayes(fit) F.stat <- FStat(fit) P.value <- pf(F.stat, df1=attr(F.stat,"df1"), df2=attr(F.stat,"df2"), lower.tail=FALSE) Gordon >all the best.
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