#!/usr/bin/env perl use strict; use warnings; #expected input: path to file, cols of counts (2 sets of 3), threshold if (!@ARGV or scalar @ARGV != 9) { print "usage snpFreq.pl /path/to/snps.txt <6 column numbers(1 based) with counts for alleles, first one group then another> #threshold outfile\n"; exit 1; } #get and verify inputs my $file = shift @ARGV; my $a1 = shift @ARGV; if ($a1 =~ /\D/ or $a1 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $a1\n"; exit 1; } my $a2 = shift @ARGV; if ($a2 =~ /\D/ or $a2 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $a2\n"; exit 1; } my $a3 = shift @ARGV; if ($a3 =~ /\D/ or $a3 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $a3\n"; exit 1; } my $b1 = shift @ARGV; if ($b1 =~ /\D/ or $b1 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $b1\n"; exit 1; } my $b2 = shift @ARGV; if ($b2 =~ /\D/ or $b2 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $b2\n"; exit 1; } my $b3 = shift @ARGV; if ($b3 =~ /\D/ or $b3 < 1) { print "Error the column number, must be an integer greater than or equal to 1. Got $b3\n"; exit 1; } my $thresh = shift @ARGV; if ($thresh !~ /^\d*\.?\d+$/) { print "Error the threshold must be a number. Got $thresh\n"; exit 1; }elsif ($thresh > .3) { print "Error the threshold can not be greater than 0.3 got $thresh\n"; exit 1; } my $outfile = shift @ARGV; #run a fishers exact test (using R) on whole table my $cmd = qq|options(warn=-1) tab <- read.table('$file', sep="\t") size <- length(tab[,1]) width <- length(tab[1,]) x <- 1:size y <- matrix(data=0, nr=size, nc=6) for(i in 1:size) { m <- matrix(c(tab[i,$a1], tab[i,$b1], tab[i,$a2], tab[i,$b2], tab[i,$a3], tab[i,$b3]), nrow=2) t <- fisher.test(m) x[i] <- t\$p.value if (x[i] >= 1) { x[i] <- .999 } n <- (tab[i,$a1] + tab[i,$a2] + tab[i,$a3] + tab[i,$b1] + tab[i,$b2] + tab[i,$b3]) n_a <- (tab[i,$a1] + tab[i,$a2] + tab[i,$a3]) y[i,1] <- ((tab[i,$a1] + tab[i,$b1])*(n_a))/n y[i,1] <- round(y[i,1],3) y[i,2] <- ((tab[i,$a2] + tab[i,$b2])*(n_a))/n y[i,2] <- round(y[i,2],3) y[i,3] <- ((tab[i,$a3] + tab[i,$b3])*(n_a))/n y[i,3] <- round(y[i,3],3) n_b <- (tab[i,$b1] + tab[i,$b2] + tab[i,$b3]) y[i,4] <- ((tab[i,$a1] + tab[i,$b1])*(n_b))/n y[i,4] <- round(y[i,4],3) y[i,5] <- ((tab[i,$a2] + tab[i,$b2])*(n_b))/n y[i,5] <- round(y[i,5],3) y[i,6] <- ((tab[i,$a3] + tab[i,$b3])*(n_b))/n y[i,6] <- round(y[i,6],3) }|; #results <- data.frame(tab[1:size,1:width], x[1:size]) #write.table(results, file="$outfile", row.names = FALSE ,col.names = FALSE,quote = FALSE, sep="\t") #q()|; my $cmd2 = qq|suppressPackageStartupMessages(library(qvalue)) qobj <- qvalue(x[1:size], lambda=seq(0,0.90,$thresh), pi0.method="bootstrap", fdr.level=0.1, robust=FALSE, smooth.log.pi0 = FALSE) q <- qobj\$qvalues results <- data.frame(tab[1:size,1:width], y[1:size,1:6], x[1:size], q[1:size]) write.table(results, file="$outfile", row.names = FALSE ,col.names = FALSE,quote = FALSE, sep="\t") q()|; #for TESTING my $pr = qq|results <- data.frame(tab[1:size,1:width], y[1:size,1:6], x[1:size]) write.table(results, file="$outfile", row.names = FALSE ,col.names = FALSE,quote = FALSE, sep="\t") q()|; open(FT, "| R --slave --vanilla") or die "Couldn't call fisher.text, $!\n"; print FT $cmd, "\n"; #fisher test print FT $cmd2, "\n"; #qvalues and results #print FT $pr, "\n"; close FT or die "Couldn't finish fisher.test, $!\n"; exit;