#!/usr/bin/env Rscript # example: # Rscript RNAseq_1_ballgown.R -o /home/yuying/rnaseqreport_test -i ./ballgown/ -l FALSE -p test # Rscript RNAseq_1_ballgown.R -o /home/yuying/rnaseqreport_test -i ./ballgown/ suppressPackageStartupMessages(library("optparse")) suppressPackageStartupMessages(library("ballgown")) # specify our desired options in a list # by default OptionParser will add an help option equivalent to # make_option(c("-h", "--help"), action="store_true", default=FALSE, # help="Show this help message and exit") option_list <- list( make_option(c("-o", "--out_dir"), type="character",default="./", help="The output directory [default ./]"), make_option(c("-i", "--input"),type="character", default=NULL, help="The directory input of expression files. It is output from ballgown software."), make_option(c("-f", "--floor_value"),metavar="number",default=0.01, help="A number to add to each value before log2 transformation to avoid infinite value.[default: 0.01]"), make_option(c("-l", "--log2_norm"), metavar="TRUE", default=TRUE, help="Perform log2 transformation on FPKM value. [default: TRUE]"), make_option(c("-p", "--project_code"), type="character",default="rnaseq", help="Project code, which is used as prefix of output file. [default: rnaseq]") ) # get command line options, if help option encountered print help and exit, # otherwise if options not found on command line then set defaults, opt <- parse_args(OptionParser(option_list=option_list)) if (is.null(opt$input)){ print_help(opt_parser) stop("At least one argument must be supplied (input file).", call.=FALSE) } #generate FPKM expression profile from ballgown outputs geballgown_expr <- ballgown(dataDir = opt$input ,samplePattern = ".*",meas = "all") expr <- gexpr(geballgown_expr) message("finish ballgown\n") #remove _1P and FPKM from colnames, _1P is from alicloud app, FPKM is added due to default output of stringtie/ballgown. nam<-colnames(expr) nam<-gsub("_1P$","",nam) nam<-gsub("^FPKM.","",nam) colnames(expr) <- nam out_dir<-paste(gsub("/$","",opt$out_dir),"/",sep="") if(opt$log2_norm==TRUE){ message("start log2 transformation\n") logexpr<-apply(expr,2,function(x){log2(x+as.numeric(opt$floor_value))}) logexpr_out<-cbind(rownames(logexpr),round(logexpr,3)) colnames(logexpr_out)[1]<-"Gene" message("Write log2 expression file\n") write.table(logexpr_out,file = paste(out_dir,opt$project_code,"_geneexp_log2FPKM.txt",sep=""),sep="\t",row.names=F,quote=F) }else{ #output expression file with fpkm expr<-cbind(rownames(expr),round(expr,3)) colnames(expr)[1]<-"Gene" message("Write fpkm expression file\n") write.table(expr,file = paste(out_dir,opt$project_code,"_geneexp_FPKM.txt",sep=""),sep="\t",row.names=F,quote=F) }