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annotate deseq2.R @ 20:89d26b11d452 draft

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planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 82fc6e1098b8af8b769ff07689704c5275b76459
author iuc
date Thu, 06 Dec 2018 15:49:22 -0500
parents c56e0689e46e
children a6fc9228e1a0
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1 #!/usr/bin/env Rscript
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3 # A command-line interface to DESeq2 for use with Galaxy
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4 # written by Bjoern Gruening and modified by Michael Love 2016.03.30
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5 #
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6 # This argument is required:
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7 #
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8 # 'factors' a JSON list object from Galaxy
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9 #
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10 # the output file has columns:
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11 #
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12 # baseMean (mean normalized count)
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13 # log2FoldChange (by default a moderated LFC estimate)
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14 # lfcSE (the standard error)
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15 # stat (the Wald statistic)
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16 # pvalue (p-value from comparison of Wald statistic to a standard Normal)
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17 # padj (adjusted p-value, Benjamini Hochberg correction on genes which pass the mean count filter)
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18 #
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19 # the first variable in 'factors' will be the primary factor.
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20 # the levels of the primary factor are used in the order of appearance in factors.
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21 #
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22 # by default, levels in the order A,B,C produces a single comparison of B vs A, to a single file 'outfile'
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23 #
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24 # for the 'many_contrasts' flag, levels in the order A,B,C produces comparisons C vs A, B vs A, C vs B,
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25 # to a number of files using the 'outfile' prefix: 'outfile.condition_C_vs_A' etc.
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26 # all plots will still be sent to a single PDF, named by the arg 'plots', with extra pages.
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27 #
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28 # fit_type is an integer valued argument, with the options from ?estimateDisperions
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29 # 1 "parametric"
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30 # 2 "local"
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31 # 3 "mean"
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33 # setup R error handling to go to stderr
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34 options( show.error.messages=F, error = function () { cat( geterrmessage(), file=stderr() ); q( "no", 1, F ) } )
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36 # we need that to not crash galaxy with an UTF8 error on German LC settings.
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37 loc <- Sys.setlocale("LC_MESSAGES", "en_US.UTF-8")
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39 library("getopt")
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40 library("tools")
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41 options(stringAsFactors = FALSE, useFancyQuotes = FALSE)
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42 args <- commandArgs(trailingOnly = TRUE)
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44 # get options, using the spec as defined by the enclosed list.
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45 # we read the options from the default: commandArgs(TRUE).
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46 spec <- matrix(c(
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47 "quiet", "q", 0, "logical",
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48 "help", "h", 0, "logical",
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49 "batch_factors", "", 1, "character",
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50 "outfile", "o", 1, "character",
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51 "countsfile", "n", 1, "character",
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52 "rlogfile", "r", 1, "character",
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53 "vstfile", "v", 1, "character",
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54 "header", "H", 0, "logical",
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55 "factors", "f", 1, "character",
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56 "files_to_labels", "l", 1, "character",
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57 "plots" , "p", 1, "character",
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58 "tximport", "i", 0, "logical",
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59 "txtype", "y", 1, "character",
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60 "tx2gene", "x", 1, "character", # a space-sep tx-to-gene map or GTF/GFF3 file
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61 "esf", "e", 1, "character",
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62 "fit_type", "t", 1, "integer",
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63 "many_contrasts", "m", 0, "logical",
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64 "outlier_replace_off" , "a", 0, "logical",
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65 "outlier_filter_off" , "b", 0, "logical",
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66 "auto_mean_filter_off", "c", 0, "logical",
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67 "beta_prior_off", "d", 0, "logical"),
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68 byrow=TRUE, ncol=4)
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69 opt <- getopt(spec)
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71 # if help was asked for print a friendly message
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72 # and exit with a non-zero error code
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73 if (!is.null(opt$help)) {
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74 cat(getopt(spec, usage=TRUE))
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75 q(status=1)
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76 }
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77
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78 # enforce the following required arguments
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79 if (is.null(opt$outfile)) {
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80 cat("'outfile' is required\n")
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81 q(status=1)
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82 }
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83 if (is.null(opt$factors)) {
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84 cat("'factors' is required\n")
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85 q(status=1)
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86 }
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87
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88 verbose <- if (is.null(opt$quiet)) {
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89 TRUE
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90 } else {
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91 FALSE
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92 }
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93
17
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94 source_local <- function(fname){
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95 argv <- commandArgs(trailingOnly = FALSE)
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96 base_dir <- dirname(substring(argv[grep("--file=", argv)], 8))
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97 source(paste(base_dir, fname, sep="/"))
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98 }
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99
17
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100 source_local('get_deseq_dataset.R')
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101
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102 suppressPackageStartupMessages({
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103 library("DESeq2")
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104 library("RColorBrewer")
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105 library("gplots")
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106 })
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107
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108 # build or read sample table
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109
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110 trim <- function (x) gsub("^\\s+|\\s+$", "", x)
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111
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112 # switch on if 'factors' was provided:
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113 library("rjson")
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114 parser <- newJSONParser()
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115 parser$addData(opt$factors)
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116 factorList <- parser$getObject()
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117 filenames_to_labels <- fromJSON(opt$files_to_labels)
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118 factors <- sapply(factorList, function(x) x[[1]])
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119 primaryFactor <- factors[1]
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120 filenamesIn <- unname(unlist(factorList[[1]][[2]]))
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121 labs = unname(unlist(filenames_to_labels[basename(filenamesIn)]))
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122 sampleTable <- data.frame(sample=basename(filenamesIn),
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123 filename=filenamesIn,
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124 row.names=filenamesIn,
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125 stringsAsFactors=FALSE)
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126 for (factor in factorList) {
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127 factorName <- trim(factor[[1]])
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128 sampleTable[[factorName]] <- character(nrow(sampleTable))
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129 lvls <- sapply(factor[[2]], function(x) names(x))
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130 for (i in seq_along(factor[[2]])) {
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131 files <- factor[[2]][[i]][[1]]
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132 sampleTable[files,factorName] <- trim(lvls[i])
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133 }
14
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134 sampleTable[[factorName]] <- factor(sampleTable[[factorName]], levels=lvls)
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135 }
14
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136 rownames(sampleTable) <- labs
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137
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138 primaryFactor <- factors[1]
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139 designFormula <- as.formula(paste("~", paste(rev(factors), collapse=" + ")))
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140
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141 # these are plots which are made once for each analysis
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142 generateGenericPlots <- function(dds, factors) {
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143 library("ggplot2")
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144 library("ggrepel")
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145 library("pheatmap")
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146
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147 rld <- rlog(dds)
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148 p <- plotPCA(rld, intgroup=rev(factors))
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149 print(p + geom_text_repel(aes_string(x = "PC1", y = "PC2", label = factor(colnames(dds))), size=3) + geom_point())
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150 dat <- assay(rld)
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151 distsRL <- dist(t(dat))
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152 mat <- as.matrix(distsRL)
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153 colors <- colorRampPalette( rev(brewer.pal(9, "Blues")) )(255)
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154 pheatmap(mat,
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155 clustering_distance_rows=distsRL,
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156 clustering_distance_cols=distsRL,
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157 col=colors,
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158 main="Sample-to-sample distances")
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159 plotDispEsts(dds, main="Dispersion estimates")
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160 }
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161
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162 # these are plots which can be made for each comparison, e.g.
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163 # once for C vs A and once for B vs A
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164 generateSpecificPlots <- function(res, threshold, title_suffix) {
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165 use <- res$baseMean > threshold
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166 if (sum(!use) == 0) {
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167 h <- hist(res$pvalue, breaks=0:50/50, plot=FALSE)
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168 barplot(height = h$counts,
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169 col = "powderblue", space = 0, xlab="p-values", ylab="frequency",
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170 main=paste("Histogram of p-values for",title_suffix))
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171 text(x = c(0, length(h$counts)), y = 0, label=paste(c(0,1)), adj=c(0.5,1.7), xpd=NA)
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172 } else {
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173 h1 <- hist(res$pvalue[!use], breaks=0:50/50, plot=FALSE)
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174 h2 <- hist(res$pvalue[use], breaks=0:50/50, plot=FALSE)
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175 colori <- c("filtered (low count)"="khaki", "not filtered"="powderblue")
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176 barplot(height = rbind(h1$counts, h2$counts), beside = FALSE,
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177 col = colori, space = 0, xlab="p-values", ylab="frequency",
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178 main=paste("Histogram of p-values for",title_suffix))
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179 text(x = c(0, length(h1$counts)), y = 0, label=paste(c(0,1)), adj=c(0.5,1.7), xpd=NA)
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180 legend("topright", fill=rev(colori), legend=rev(names(colori)), bg="white")
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181 }
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182 plotMA(res, main= paste("MA-plot for",title_suffix), ylim=range(res$log2FoldChange, na.rm=TRUE))
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183 }
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184
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185 if (verbose) {
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186 cat(paste("primary factor:",primaryFactor,"\n"))
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187 if (length(factors) > 1) {
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188 cat(paste("other factors in design:",paste(factors[-length(factors)],collapse=","),"\n"))
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189 }
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190 cat("\n---------------------\n")
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191 }
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192
17
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193 dds <- get_deseq_dataset(sampleTable, header=opt$header, designFormula=designFormula, tximport=opt$tximport, txtype=opt$txtype, tx2gene=opt$tx2gene)
18
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194 # estimate size factors for the chosen method
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195 if(!is.null(opt$esf)){
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196 dds <- estimateSizeFactors(dds, type=opt$esf)
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197 }
17
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198 apply_batch_factors <- function (dds, batch_factors) {
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199 rownames(batch_factors) <- batch_factors$identifier
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200 batch_factors <- subset(batch_factors, select = -c(identifier, condition))
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201 dds_samples <- colnames(dds)
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202 batch_samples <- rownames(batch_factors)
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203 if (!setequal(batch_samples, dds_samples)) {
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204 stop("Batch factor names don't correspond to input sample names, check input files")
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205 }
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206 dds_data <- colData(dds)
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207 # Merge dds_data with batch_factors using indexes, which are sample names
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208 # Set sort to False, which maintains the order in dds_data
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209 reordered_batch <- merge(dds_data, batch_factors, by.x = 0, by.y = 0, sort=F)
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210 batch_factors <- reordered_batch[, ncol(dds_data):ncol(reordered_batch)]
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211 for (factor in colnames(batch_factors)) {
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212 dds[[factor]] <- batch_factors[[factor]]
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213 }
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214 colnames(dds) <- reordered_batch[,1]
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215 return(dds)
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216 }
15
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217
17
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218 if (!is.null(opt$batch_factors)) {
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219 batch_factors <- read.table(opt$batch_factors, sep="\t", header=T)
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220 dds <- apply_batch_factors(dds = dds, batch_factors = batch_factors)
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221 batch_design <- colnames(batch_factors)[-c(1,2)]
d9e5cadc7f0b planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit b95582cea8320d5488056a9576474f79cec53be8
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222 designFormula <- as.formula(paste("~", paste(c(batch_design, rev(factors)), collapse=" + ")))
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223 design(dds) <- designFormula
6
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224 }
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225
15
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226 if (verbose) {
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227 cat("DESeq2 run information\n\n")
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228 cat("sample table:\n")
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229 print(sampleTable[,-c(1:2),drop=FALSE])
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230 cat("\ndesign formula:\n")
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231 print(designFormula)
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232 cat("\n\n")
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233 cat(paste(ncol(dds), "samples with counts over", nrow(dds), "genes\n"))
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234 }
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235
6
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236 # optional outlier behavior
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237 if (is.null(opt$outlier_replace_off)) {
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238 minRep <- 7
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239 } else {
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240 minRep <- Inf
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241 if (verbose) cat("outlier replacement off\n")
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242 }
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243 if (is.null(opt$outlier_filter_off)) {
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244 cooksCutoff <- TRUE
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245 } else {
6
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246 cooksCutoff <- FALSE
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247 if (verbose) cat("outlier filtering off\n")
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248 }
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249
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250 # optional automatic mean filtering
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251 if (is.null(opt$auto_mean_filter_off)) {
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252 independentFiltering <- TRUE
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253 } else {
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254 independentFiltering <- FALSE
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255 if (verbose) cat("automatic filtering on the mean off\n")
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256 }
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257
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258 # shrinkage of LFCs
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259 if (is.null(opt$beta_prior_off)) {
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260 betaPrior <- TRUE
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261 } else {
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262 betaPrior <- FALSE
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263 if (verbose) cat("beta prior off\n")
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264 }
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265
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266 # dispersion fit type
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267 if (is.null(opt$fit_type)) {
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268 fitType <- "parametric"
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269 } else {
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270 fitType <- c("parametric","local","mean")[opt$fit_type]
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271 }
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272
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273 if (verbose) cat(paste("using disperion fit type:",fitType,"\n"))
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parents:
diff changeset
274
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
275 # run the analysis
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
276 dds <- DESeq(dds, fitType=fitType, betaPrior=betaPrior, minReplicatesForReplace=minRep)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
277
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
278 # create the generic plots and leave the device open
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
279 if (!is.null(opt$plots)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
280 if (verbose) cat("creating plots\n")
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
281 pdf(opt$plots)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
282 generateGenericPlots(dds, factors)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
283 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
284
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
285 n <- nlevels(colData(dds)[[primaryFactor]])
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
286 allLevels <- levels(colData(dds)[[primaryFactor]])
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
287
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
288 if (!is.null(opt$countsfile)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
289 normalizedCounts<-counts(dds,normalized=TRUE)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
290 write.table(normalizedCounts, file=opt$countsfile, sep="\t", col.names=NA, quote=FALSE)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
291 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
292
18
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
293 if (!is.null(opt$rlogfile)) {
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
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diff changeset
294 rLogNormalized <-rlogTransformation(dds)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
295 rLogNormalizedMat <- assay(rLogNormalized)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
296 write.table(rLogNormalizedMat, file=opt$rlogfile, sep="\t", col.names=NA, quote=FALSE)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
297 }
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
298
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
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diff changeset
299 if (!is.null(opt$vstfile)) {
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
300 vstNormalized<-varianceStabilizingTransformation(dds)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
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diff changeset
301 vstNormalizedMat <- assay(vstNormalized)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
302 write.table(vstNormalizedMat, file=opt$vstfile, sep="\t", col.names=NA, quote=FALSE)
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
303 }
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
304
3bf1b3ec1ddf planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 448dccb0c02aba00d8301247b0f0f406ab3d4fa2
iuc
parents: 17
diff changeset
305
6
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
306 if (is.null(opt$many_contrasts)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
307 # only contrast the first and second level of the primary factor
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
308 ref <- allLevels[1]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
309 lvl <- allLevels[2]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
310 res <- results(dds, contrast=c(primaryFactor, lvl, ref),
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
311 cooksCutoff=cooksCutoff,
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
312 independentFiltering=independentFiltering)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
313 if (verbose) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
314 cat("summary of results\n")
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
315 cat(paste0(primaryFactor,": ",lvl," vs ",ref,"\n"))
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
316 print(summary(res))
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
317 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
318 resSorted <- res[order(res$padj),]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
319 outDF <- as.data.frame(resSorted)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
320 outDF$geneID <- rownames(outDF)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
321 outDF <- outDF[,c("geneID", "baseMean", "log2FoldChange", "lfcSE", "stat", "pvalue", "padj")]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
322 filename <- opt$outfile
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
323 write.table(outDF, file=filename, sep="\t", quote=FALSE, row.names=FALSE, col.names=FALSE)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
324 if (independentFiltering) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
325 threshold <- unname(attr(res, "filterThreshold"))
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
326 } else {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
327 threshold <- 0
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
328 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
329 title_suffix <- paste0(primaryFactor,": ",lvl," vs ",ref)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
330 if (!is.null(opt$plots)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
331 generateSpecificPlots(res, threshold, title_suffix)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
332 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
333 } else {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
334 # rotate through the possible contrasts of the primary factor
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
335 # write out a sorted table of results with the contrast as a suffix
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
336 # add contrast specific plots to the device
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
337 for (i in seq_len(n-1)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
338 ref <- allLevels[i]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
339 contrastLevels <- allLevels[(i+1):n]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
340 for (lvl in contrastLevels) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
341 res <- results(dds, contrast=c(primaryFactor, lvl, ref),
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
342 cooksCutoff=cooksCutoff,
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
343 independentFiltering=independentFiltering)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
344 resSorted <- res[order(res$padj),]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
345 outDF <- as.data.frame(resSorted)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
346 outDF$geneID <- rownames(outDF)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
347 outDF <- outDF[,c("geneID", "baseMean", "log2FoldChange", "lfcSE", "stat", "pvalue", "padj")]
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
348 filename <- paste0(opt$outfile,".",primaryFactor,"_",lvl,"_vs_",ref)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
349 write.table(outDF, file=filename, sep="\t", quote=FALSE, row.names=FALSE, col.names=FALSE)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
350 if (independentFiltering) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
351 threshold <- unname(attr(res, "filterThreshold"))
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
352 } else {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
353 threshold <- 0
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
354 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
355 title_suffix <- paste0(primaryFactor,": ",lvl," vs ",ref)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
356 if (!is.null(opt$plots)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
357 generateSpecificPlots(res, threshold, title_suffix)
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
358 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
359 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
360 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
361 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
362
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
363 # close the plot device
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
364 if (!is.null(opt$plots)) {
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
365 cat("closing plot device\n")
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
diff changeset
366 dev.off()
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
iuc
parents:
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367 }
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
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4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
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369 cat("Session information:\n\n")
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
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parents:
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4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
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parents:
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371 sessionInfo()
4939397c4706 planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/deseq2 commit 3bc8d91ee546682ef8e9303bd1044bb14cf21b07
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