# HG changeset patch # User artbio # Date 1569509455 14400 # Node ID 252eded618481a609f8c9049c5ce124e1132efaf "planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/gsc_scran_normalize commit ddcf915dd9b690d7f3876e08b939adde36cbb8dd" diff -r 000000000000 -r 252eded61848 scran-normalize.R --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/scran-normalize.R Thu Sep 26 10:50:55 2019 -0400 @@ -0,0 +1,91 @@ +# load packages that are provided in the conda env +options( show.error.messages=F, + error = function () { cat( geterrmessage(), file=stderr() ); q( "no", 1, F ) } ) +loc <- Sys.setlocale("LC_MESSAGES", "en_US.UTF-8") +warnings() + +library(optparse) +library(scran) + +#Arguments +option_list = list( + make_option( + c("-d", "--data"), + default = NA, + type = 'character', + help = "Input file that contains count values to transform" + ), + make_option( + c("-s", "--sep"), + default = '\t', + type = 'character', + help = "File separator [default : '%default' ]" + ), + make_option( + "--cluster", + default=FALSE, + action="store_true", + type = 'logical', + help = "Whether to calculate the size factor per cluster or on all cell" + ), + make_option( + c("-m", "--method"), + default = 'hclust', + type = 'character', + help = "The clustering method to use for grouping cells into cluster : hclust or igraph [default : '%default' ]" + ), + make_option( + "--size", + default = 100, + type = 'integer', + help = "Minimal number of cells in each cluster : hclust or igraph [default : '%default' ]" + ), + make_option( + c("-o", "--out"), + default = "res.tab", + type = 'character', + help = "Output name [default : '%default' ]" + ) +) + +opt = parse_args(OptionParser(option_list = option_list), + args = commandArgs(trailingOnly = TRUE)) + +if (opt$sep == "tab") {opt$sep = "\t"} + +data = read.table( + opt$data, + check.names = FALSE, + header = TRUE, + row.names = 1, + sep = opt$sep +) + +## Import data as a SingleCellExperiment object +sce <- SingleCellExperiment(list(counts=as.matrix(data))) + + +if(opt$cluster){ + clusters <- quickCluster(sce, min.size = opt$size, method = opt$method) + + ## Compute sum factors + sce <- computeSumFactors(sce, cluster = clusters) +} else { + + ## Compute sum factors + sce <- computeSumFactors(sce) +} + +sce <- normalize(sce) + +logcounts <- data.frame(genes = rownames(sce), round(logcounts(sce), digits=5), check.names = F) + + +write.table( + logcounts, + opt$out, + col.names = T, + row.names = F, + quote = F, + sep = "\t" +) diff -r 000000000000 -r 252eded61848 scran_normalize.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/scran_normalize.xml Thu Sep 26 10:50:55 2019 -0400 @@ -0,0 +1,107 @@ + + Normalize raw counts expression values using deconvolution size factors + + r-optparse + bioconductor-scran + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +**What it does** + +Takes a raw count expression matrix and returns a table of log transformed scran-normalized expression values. + +This computes size factors that are used to scale the counts in each cell. The assumption is that +most genes are not differentially expressed (DE) between cells, such that any differences in +expression across the majority of genes represents some technical bias that should be removed. + +Cell-specific biases are normalized using the computeSumFactors method, which implements the +deconvolution strategy for scaling normalization (A. T. Lun, Bach, and Marioni 2016). It creates a reference : + - if no clustering step : the average count of all transcriptomes + - if you choose to cluster your cells : the average count of each cluster. +Then it pools cells and then sum their expression profiles. The size factor is described as the median ration +between the count sums and the average across all genes. Finally it constructs a linear distribution (deconvolution method) +of size factors by taking multiple pools of cells. + +You can apply this method on cell cluster instead of your all set of cells by using quickCluster. +It defines cluster using distances based on Spearman correlation on counts between cells, there is two available methods : + + - *hclust* : hierarchical clustering on the distance matrix and dynamic tree cut. + - *igraph* : constructs a Shared Nearest Neighbor graph (SNN) on the distance matrix and identifies highly connected communities. + + +Note: First header row must NOT start with a '#' comment character + + + + + @Article{, + author = {Aaron T. L. Lun and Davis J. McCarthy and John C. Marioni}, + title = {A step-by-step workflow for low-level analysis of single-cell RNA-seq data with Bioconductor}, + journal = {F1000Res.}, + year = {2016}, + volume = {5}, + pages = {2122}, + doi = {10.12688/f1000research.9501.2}, + } + + + diff -r 000000000000 -r 252eded61848 test-data/counts.tab --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/counts.tab Thu Sep 26 10:50:55 2019 -0400 @@ -0,0 +1,101 @@ +Genes 1001000174.B1 1001000177.C5 1001000182.H4 1001000175.A2 1001000230.B11 1001000237.E9 1001000173.E10 1001000241.E7 1001000249.A3 1001000247.F11 1001000240.D9 1001000178.G5 1001000249.E3 1001000174.H6 1001000181.F8 1001000177.A10 1001000235.D2 1001000248.E7 1001000012.B6 1001000179.H4 1001000230.C1 1001000241.G11 1001000174.G2 1001000252.C6 1001000186.D11 1001000174.A10 1001000255.E9 1001000242.B5 1001000179.F3 1001000185.F9 1001000267.F8 1001000183.G10 1001000247.E7 1001000031.A2 1001000271.B1 1001000187.G6 1001000236.C6 1001000187.D6 1001000235.E10 1001000036.C1 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