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Perl script for chi-squared test !

  • Public
By Neelam Jha 412 days ago
#!/usr/bin/perl # # chidi.pl # # A script to perform a chi-squared test of the dinucleotide frequencies of two FASTA files # Last updated by: $Author$ # Last updated on: $Date$ use strict; use warnings; use Getopt::Long; use FAlite; # sanity checks die "Usage: chidi.pl <file1> <file2>\n" if (!$ARGV[1]); my @dinucs = qw (AA AC AG AT CA CC CG CT GA GC GG GT TA TC TG TT); # hashes for obersered and expected dinucleotide frequencies of both files my %file1_ob; my %file2_ob; my %file1_ex; my %file2_ex; ############################################################ # Read sequence file 1 ############################################################ open(FILE,"$ARGV[0]") || die "Can't open $ARGV[0]\n"; my $fasta = new FAlite(\*FILE); # loop through each sequence in file 1 while(my $entry = $fasta->nextEntry) { my $seq = uc($entry->seq); # to count dinucleotides, loop through sequence, take 2 bp and increment the hash counter foreach my $i (0..length($seq)){ my $tmp = substr($seq,$i,2); $file1_ob{$tmp}++; } } close(FILE); ############################################################ # Read sequence file 2 ############################################################ open(FILE,"$ARGV[1]") || die "Can't open $ARGV[1]\n"; $fasta = new FAlite(\*FILE); # loop through each sequence in file 1 while(my $entry = $fasta->nextEntry) { my $seq = uc($entry->seq); # to count dinucleotides, loop through sequence, take 2 bp and increment the hash counter foreach my $i (0..length($seq)){ my $tmp = substr($seq,$i,2); $file2_ob{$tmp}++; } } close(FILE); ############################################################ # Perform chi-squared test ############################################################ # need total of all counts in both sequences, plus totals of 'rows' in chi-square table my $total; my $row1; my $row2; foreach my $di (@dinucs){ $row1 += $file1_ob{$di}; $row2 += $file2_ob{$di}; $total += ($file1_ob{$di} + $file2_ob{$di}); } # now calculate expected values foreach my $di (@dinucs){ # calculate (column total * row total) / $total $file1_ex{$di} = sprintf("%.2f",(($file1_ob{$di}+$file2_ob{$di}) * $row1) / $total); $file2_ex{$di} = sprintf("%.2f",(($file1_ob{$di}+$file2_ob{$di}) * $row2) / $total); } # now calculate chi-squared values my ($chi1,$chi2); my $chi_total; print "\tObs1\tExp2\t\tChi1\tObs2\tExp2\t\tChi2\n"; foreach my $di (@dinucs){ $chi1 = sprintf("%.2f",(($file1_ob{$di} - $file1_ex{$di})**2)/$file1_ex{$di}); $chi2 = sprintf("%.2f",(($file2_ob{$di} - $file2_ex{$di})**2)/$file2_ex{$di}); print "$di\t$file1_ob{$di}\t$file1_ex{$di}\t$chi1\t$file2_ob{$di}\t$file2_ex{$di}\t$chi2\n"; $chi_total += ($chi1+$chi2); } printf "Chi squared value = %6.2f\n", $chi_total; print "Significance level at 5% = 25.00\n"; print "Significance level at 1% = 30.58\n"; exit(0);