#!/usr/bin/env perl # # Copyright (C) 2009 Genome Research Ltd. # # Author: Heng Li # # Permission is hereby granted, free of charge, to any person obtaining a copy # of this software and associated documentation files (the "Software"), to deal # in the Software without restriction, including without limitation the rights # to use, copy, modify, merge, publish, distribute, sublicense, and/or sell # copies of the Software, and to permit persons to whom the Software is # furnished to do so, subject to the following conditions: # # The above copyright notice and this permission notice shall be included in # all copies or substantial portions of the Software. # # THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR # IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, # FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL # THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER # LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING # FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER # DEALINGS IN THE SOFTWARE. use strict; use warnings; use Getopt::Std; &bowtie2sam; exit; sub bowtie2sam { my %opts = (); die("Usage: bowtie2sam.pl \n") if (@ARGV == 0 && -t STDIN); # core loop my (@s, $last, @staging, $k, $best_s, $subbest_s, $best_k); $last = ''; while (<>) { my ($name, $nm) = &bowtie2sam_aux($_, \@s); # read_name, number of mismatches if ($name eq $last) { # I do not know whether the multiple hits are ordered on the # number of mismatches. I assume they are not and so I have to # keep all these multiple hits in memory. @{$staging[$k]} = @s; if ($best_s > $nm) { $subbest_s = $best_s; $best_s = $nm; $best_k = $k; } elsif ($subbest_s > $nm) { $subbest_s = $nm; } ++$k; } else { if ($last) { if ($best_s == $subbest_s) { $staging[$best_k][4] = 0; } elsif ($subbest_s - $best_s == 1) { $staging[$best_k][4] = 15 if ($staging[$best_k][4] > 15); } print join("\t", @{$staging[$best_k]}), "\n"; } $k = 1; $best_s = $nm; $subbest_s = 1000; $best_k = 0; @{$staging[0]} = @s; $last = $name; } } print join("\t", @{$staging[$best_k]}), "\n" if ($best_k >= 0); } sub bowtie2sam_aux { my ($line, $s) = @_; chomp($line); my @t = split("\t", $line); my $ret; @$s = (); # read name $s->[0] = $ret = $t[0]; $s->[0] =~ s/\/[12]$//g; # initial flag (will be updated later) $s->[1] = 0; # read & quality $s->[9] = $t[4]; $s->[10] = $t[5]; # cigar $s->[5] = length($s->[9]) . "M"; # coor $s->[2] = $t[2]; $s->[3] = $t[3] + 1; $s->[1] |= 0x10 if ($t[1] eq '-'); # mapQ $s->[4] = $t[6] == 0? 25 : 0; # mate coordinate $s->[6] = '*'; $s->[7] = $s->[8] = 0; # aux my $nm = @t - 7; push(@$s, "NM:i:" . (@t-7)); push(@$s, "X$nm:i:" . ($t[6]+1)); my $md = ''; if ($t[7]) { $_ = $t[7]; my $a = 0; while (/(\d+):[ACGTN]>([ACGTN])/gi) { my ($y, $z) = ($1, $2); $md .= (int($y)-$a) . $z; $a += $y - $a + 1; } $md .= length($s->[9]) - $a; } else { $md = length($s->[9]); } push(@$s, "MD:Z:$md"); return ($ret, $nm); }