## ----echo=FALSE, results="hide", message=FALSE-------------------------------- require(knitr) opts_chunk$set(error=FALSE, message=FALSE, warning=FALSE) ## ----setup, echo=FALSE, message=FALSE----------------------------------------- library(scran) library(BiocParallel) set.seed(100) self <- BiocStyle::Biocpkg("scran") ## ----------------------------------------------------------------------------- library(scran) library(scRNAseq) sce <- GrunPancreasData() sce # Quickly analyzing it to generate bits and pieces that we can # reuse in the rest of this vignette. library(scrapper) results <- analyze.se(sce, num.threads=2) sce.ready <- results$x ## ----------------------------------------------------------------------------- library(scran) clusters <- quickCluster(sce.ready) sce.normed <- computeSumFactors(sce.ready, clusters=clusters) summary(sizeFactors(sce.normed)) ## ----------------------------------------------------------------------------- subout <- quickSubCluster(sce.ready, groups=clusters) table(metadata(subout)$subcluster) # formatted as '.' ## ----------------------------------------------------------------------------- var.explained <- metadata(sce.ready)$PCA$variance.explained stats <- rowData(sce.ready) total.tech.var.in.hvgs <- sum(stats[stats$hvg,"fitted"]) total.var.in.hvgs <- sum(stats[stats$hvg,"variances"]) chosen <- denoisePCANumber(var.explained, total.tech.var.in.hvgs, total.var.in.hvgs) chosen ## ----------------------------------------------------------------------------- reducedDim(sce.ready, "PCA.denoised") <- reducedDim(sce.ready, "PCA")[,1:chosen] ## ----------------------------------------------------------------------------- # Using the first 200 HVs, which are the most interesting anyway. of.interest <- order(rowData(sce.ready)$residuals, decreasing=TRUE)[1:200] cor.pairs <- correlatePairs(sce.ready, subset.row=of.interest) cor.pairs ## ----------------------------------------------------------------------------- cor.pairs2 <- correlatePairs(sce.ready, subset.row=of.interest, block=sce.ready$donor) ## ----------------------------------------------------------------------------- cor.genes <- correlateGenes(cor.pairs) cor.genes ## ----------------------------------------------------------------------------- sessionInfo()