## ----options, include=FALSE, echo=FALSE------------------------------------ library(BiocStyle) knitr::opts_chunk$set(warning=FALSE, error=FALSE, message=FALSE) ## ----construct------------------------------------------------------------- library(SingleCellExperiment) counts <- matrix(rpois(100, lambda = 10), ncol=10, nrow=10) sce <- SingleCellExperiment(assays = list(counts = counts)) sce ## ----coerce---------------------------------------------------------------- se <- SummarizedExperiment(list(counts=counts)) as(se, "SingleCellExperiment") ## ----fluidigm-------------------------------------------------------------- library(scRNAseq) sce <- ReprocessedAllenData("tophat_counts") sce ## ----subset---------------------------------------------------------------- counts <- assay(sce, "tophat_counts") libsizes <- colSums(counts) size.factors <- libsizes/mean(libsizes) logcounts(sce) <- log2(t(t(counts)/size.factors) + 1) assayNames(sce) ## ----pca------------------------------------------------------------------- pca_data <- prcomp(t(logcounts(sce)), rank=50) library(Rtsne) set.seed(5252) tsne_data <- Rtsne(pca_data$x[,1:50], pca = FALSE) reducedDims(sce) <- list(PCA=pca_data$x, TSNE=tsne_data$Y) sce ## -------------------------------------------------------------------------- reducedDims(sce) reducedDimNames(sce) head(reducedDim(sce, "PCA")[,1:2]) head(reducedDim(sce, "TSNE")[,1:2]) ## -------------------------------------------------------------------------- dim(reducedDim(sce, "PCA")) dim(reducedDim(sce[,1:10], "PCA")) ## -------------------------------------------------------------------------- counts(sce) <- assay(sce, "tophat_counts") sce dim(counts(sce)) ## -------------------------------------------------------------------------- is.spike <- grepl("^ERCC-", rownames(sce)) sce <- splitAltExps(sce, ifelse(is.spike, "ERCC", "gene")) altExpNames(sce) ## -------------------------------------------------------------------------- altExp(sce) ## -------------------------------------------------------------------------- rowData(altExp(sce))$concentration <- runif(nrow(altExp(sce))) rowData(altExp(sce)) rowData(sce) ## -------------------------------------------------------------------------- sessionInfo()