# Don't delete this chunk if you are using the mosaic package # This loads the mosaic and dplyr packages require(mosaic)
# Some customization. You can alter or delete as desired (if you know what you are doing). # This changes the default colors in lattice plots. trellis.par.set(theme=theme.mosaic()) # knitr settings to control how R chunks work. require(knitr) opts_chunk$set( tidy=FALSE, # display code as typed size="small" # slightly smaller font for code )
# Load additional packages here. Uncomment the line below to use Project MOSAIC data sets. # require(mosaicData)
library(devtools) install_github("jtlovell/physGenomicsPVFinal") library(physGenomicsPVFinal)
library(physGenomicsPVFinal) data(temple2012_6treatments) ### for conviencence, rename info and counts info<-info12 counts<-counts12 ################################# # Part 1: Full model with Treatment ################################# ## Run LIMMA, with subplot treated as repeated measures of the same plant. ### The only response is treatment. Sub_Block is the sub-block in the split plot design, containing two AP13 ### plants. The duplicate correlation and blocking in LIMMA should account for correlations between these plants ### We do not estimate intercept, so that the model only looks at the effects of Treatment, and the F-test reflects this ## Run the modeling pipeline stats<-pipeLIMMA(counts=counts, info=info, block=info$Sub_Block, formula="~ Treatment") ### Extract voom-normalized counts v<-stats$voom[["E"]] ### Extract statistics ### Here, the F statistics = "moderated F-statistics for testing all contrasts defined by the columns ### of fit simultaneously equal to zero" ### See LIMMA::ebayes for descriptions of other statistics stats.fullmodel<-stats$simpleStats ### Fstatistics for the whole model stats.allests<-stats$stats ################################# # Part 2: All pairwise contrasts ################################# ## Generate a design matrix that specifies all relavant contrasts f<-info$Treatment design <- model.matrix(~0+f) contrast.matrix<-makeContrasts(f25th-flow, fmean-flow, fambient-flow, f75th-flow, fhigh-flow, fmean-f25th, fambient-f25th, f75th-f25th, fhigh-f25th, fambient-fmean, f75th-fmean, fhigh-fmean, f75th-fambient, fhigh-fambient, fhigh-f75th, levels=design) ## run the LIMMA contrasts pipeline ## This returns a simple dataset with relevant statistics for each contrast, plus overall F-test. lim.contrasts<-anovaLIMMA(counts=counts, design=design, block=info$Sub_Block, contrast.matrix=contrast.matrix) ################################# # Part 3: Run PCA ################################# pca<-voom2PCA(v=v, info=info, ids=info$ID) ggplot(pca, aes(x=PC1, y=PC2, col=Treatment, alpha=Treatment))+ theme_bw()+geom_point(size=4) ################################# # Part 4: Subset to lines with physiology data ################################# phys.lines<-info$ID[!is.na(info$order)] counts<-counts[,phys.lines] info<-info[info$ID %in% phys.lines,] info$Treatment<-factor(info$Treatment, levels=c("low","mean","high")) ################################# # Part 5: Run model with MDWP as the predictor ################################# stats<-pipeLIMMA(counts=counts, info=info, block=info$Sub_Block, formula="~ MDWP") stats.fullmodel.mdwp<-stats$simpleStats stats.allests.mdwp<-stats$stats ################################# # Part 6: Run model with MDWP as the predictor ################################# stats<-pipeLIMMA(counts=counts, info=info, block=info$Sub_Block, formula="~ order") stats.fullmodel.order<-stats$simpleStats stats.allests.order<-stats$stats
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