view scripts/dropletutils.Rscript @ 1:cfe1e6c28d95 draft

"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/dropletutils/ commit 40d4a92c62d75fe931baba8657cde006a26d84cf"

## Load in data
args = commandArgs(trailingOnly = T)
if (length(args) != 1){
    stop("Please provide the config file")
}

suppressWarnings(suppressPackageStartupMessages(require(DropletUtils)))
suppressWarnings(suppressPackageStartupMessages(require(Matrix)))
suppressWarnings(suppressPackageStartupMessages(require(scater)))

source(args[1])

## Helper functions
setSparse <- function(obj){
    return(as(obj, "dgCMatrix"))
}

writeTSV <- function(fileout, obj){
    write.table(as.matrix(obj), file=fileout, col.names=NA, sep='\t', quote=FALSE)
}

writeOut <- function(counts, fileout, typeout){
    if (typeout == "tsv"){
        writeTSV(fileout, counts)
    }
    else if (typeout == "h5ad"){
        write10xCounts(fileout, counts, type="HDF5", overwrite=TRUE)
    }
    else if (typeout == "directory"){
        write10xCounts(fileout, counts, type="sparse", overwrite=TRUE)
    }
}


read10xFiles <- function(filein, typein){
    sce <- NULL
    if (typein == "tsv"){
        ## Exploding memory problems occured here
        ## - solution is to use the readSparseCounts function from scater
        sce <- SingleCellExperiment(assays = list(counts = readSparseCounts(filein)))
    }
    else if (typein == "h5ad"){
        sce <- read10xCounts(filein, col.names=T, type="HDF5")   # use barcodes.tsv as column names
    }
    else if (typein == "directory"){
        sce <- read10xCounts(filein, col.names=T, type="sparse")
    }
    counts(sce) <- setSparse(counts(sce))
    return(sce)
}


## Methods


doEmptyDrops <- function(files, eparams, in.type="directory", out.type="h5ad", fdr_threshold = 0.01){
    sce <- read10xFiles(files$infile, in.type)

    eparams$... <- NULL ## hack
    eparams$m = Matrix(counts(sce), sparse=TRUE)
    
    e.out <- do.call(emptyDrops, c(eparams))
    
    bar.names <- colnames(sce)
    if (length(bar.names) != nrow(e.out)){    
        stop("Length of barcodes and output metrics don't match.")
    }
    e.out <- cbind(bar.names, e.out)
    e.out$is.Cell <- e.out$FDR <= fdr_threshold
    e.out$is.CellAndLimited <- e.out$is.Cell & e.out$Limited
    
    # Write to table
    writeTSV(files$table, e.out)
    
    # Print to log
    print(table(Limited=e.out$Limited, Significant=e.out$is.Cell))
    
    # Write to Plot
    png(files$plot)
    plot(e.out$Total, -e.out$LogProb, col=ifelse(e.out$is.Cell, "red", "black"),
    xlab="Total UMI count", ylab="-Log Probability")
    dev.off()
    
    # Filtered
    called <- e.out$is.CellAndLimited
    called[is.na(called)] <- FALSE    # replace NA's with FALSE
    sce.filtered <- sce[,called]
    
    writeOut(counts(sce.filtered), files$out, out.type)
}


doDefaultDrops <- function(files, dparams, in.type="directory", out.type="h5ad"){
    sce <- read10xFiles(files$infile, in.type)

    dparams$m = counts(sce)
    called <- do.call(defaultDrops, c(dparams))
    print(table(called))
        
    # Filtered
    sce.filtered <- sce[,called]
    
    writeOut(Matrix(counts(sce.filtered),sparse=TRUE), files$out, out.type)
}


doBarcodeRankings <- function(files, bparams, in.type="directory"){
    sce <- read10xFiles(files$infile, in.type)

    bparams$... <- NULL ## hack
    bparams$m = counts(sce)

    br.out <- do.call(barcodeRanks, c(bparams))
    
    png(files$plot)
    plot(br.out$rank, br.out$total, log="xy", xlab="(log) Rank", ylab="(log) Total Number of Barcodes")
    o <- order(br.out$rank)
    lines(br.out$rank[o], br.out$fitted[o], col="red")

    abline(h=br.out$knee, col="dodgerblue", lty=2)
    abline(h=br.out$inflection, col="forestgreen", lty=2)
    legend("bottomleft", lty=2, col=c("dodgerblue", "forestgreen"), legend=c("knee", "inflection"))
    dev.off()
    
    print(paste("knee =", br.out$knee, ", inflection = ", br.out$inflection))
}

## Main
set.seed(seed.val)

if (do.method == "barcodeRankings") {
    doBarcodeRankings(files, bparams, in.type)

} else if (do.method == "defaultDrops") {
    doDefaultDrops(files, dparams, in.type, out.type)

} else if (do.method == "emptyDrops") {
    doEmptyDrops(files, eparams, in.type, out.type, empty.fdr_threshold)
}