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#!/usr/bin/env perl
#
# Copyright 2011, Julian Catchen <jcatchen@uoregon.edu>
#
# This file is part of Stacks.
#
# Stacks is free software: you can redistribute it and/or modify
# it under the terms of the GNU General Public License as published by
# the Free Software Foundation, either version 3 of the License, or
# (at your option) any later version.
#
# Stacks is distributed in the hope that it will be useful,
# but WITHOUT ANY WARRANTY; without even the implied warranty of
# MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
# GNU General Public License for more details.
#
# You should have received a copy of the GNU General Public License
# along with Stacks. If not, see <http://www.gnu.org/licenses/>.
#
use strict;
use constant stacks_version => "_VERSION_";
my $debug = 0;
my $paired = 0;
my $amos = 0;
my $ins_len_dist = 0;
my $single_path = "";
my $paired_path = "";
my $sanger_path = "";
my $out_path = ".";
my $white_list = "";
my $hash_len = 27;
my $insert_len = 0;
my $exp_cov = 0;
my $cov_cut = 0.0;
my $min_len = 0;
my $read_trk = 0;
my $clean = 1;
my $collate = 0;
my $exe_path = "";
my $velveth = "velveth";
my $velvetg = "velvetg";
parse_command_line();
if (length($exe_path) > 0) {
$velveth = $exe_path . "/" . $velveth;
$velvetg = $exe_path . "/" . $velvetg;
}
#
# Test that we can execute the velvet programs
#
die ("Unable to find '" . $velveth . "'.\n") if (!-e $velveth || !-x $velveth);
die ("Unable to find '" . $velvetg . "'.\n") if (!-e $velvetg || !-x $velvetg);
my (@locus_files, $num_files, $file, $input_file, $output_file, $hres_file, $gres_file, $collate_fh);
build_file_list(\@locus_files);
$num_files = scalar(@locus_files);
if ($collate) {
open($collate_fh, ">$out_path/collated.fa") or die("Unable to open collate file, $!\n");
}
my ($sing_data, $pair_data, $sang_data, $ins, $cov, $afg, $cut, $min, $read, $cln);
$ins = $paired > 0 ? "-ins_length $insert_len" : "-ins_length auto";
$cov = $paired > 0 ? "-exp_cov $exp_cov" : "-exp_cov auto";
$cut = $cov_cut > 0 ? "-cov_cutoff $cov_cut" : "-cov_cutoff auto";
$read = $read_trk > 0 ? "-read_trkg yes" : "";
$cln = $clean > 0 ? "-very_clean yes" : "";
#$min = $min_len > 0 ? "-min_contig_lgth $min_len" : ""
#
# Write out the parameters for this assembly
#
open(PARAM, "> $out_path/velvet_parameters.txt") or die("Unable to open parameter file: $!\n");
print PARAM
"Single-end Path: ", $single_path, "\n",
"Paired-end Path: ", $paired_path, "\n",
"Sanger Path: ", $sanger_path, "\n",
"Hash Length: ", $hash_len, "\n",
"Insert Length: ", $ins, "\n",
"Coverage: ", $cov, "\n",
"Coverage Cutoff: ", $cut, "\n",
"Miniumum contig length: ", $min, "\n",
"Read tracking: ", $read, "\n",
"Very Clean: ", $cln, "\n";
close(PARAM);
my $i = 1;
foreach $file (@locus_files) {
($file) = ($file =~ /(.+)\.fa/);
$output_file = $out_path . "/" . $file;
$hres_file = $out_path . "/" . $file . "-h.output";
$gres_file = $out_path . "/" . $file . "-g.output";
$sing_data = length($single_path) > 0 ? '-short ' . $single_path . "/" . $file . ".fa" : "";
$pair_data = length($paired_path) > 0 ? '-shortPaired ' . $paired_path . "/" . $file . ".fa" : "";
$sang_data = length($sanger_path) > 0 ? '-long ' . $sanger_path . "/" . $file . ".fa" : "";
print STDERR "Assembling locus '$file'; run $i of $num_files \r";
# Execute velveth to build hash table, then velvetg to assemble
print STDERR "$velveth $output_file $hash_len -fasta $sing_data $pair_data $sang_data &> $hres_file\n" if ($debug);
`$velveth $output_file $hash_len -fasta $sing_data $pair_data $sang_data &> $hres_file`;
print STDERR "$velvetg $output_file $ins $cov $cut $min $read $cln &> $gres_file\n" if ($debug);
`$velvetg $output_file $ins $cov $cut $min $read $cln &> $gres_file`;
collate_and_clean($out_path, $file, $collate_fh) if ($collate);
$i++;
}
close($collate_fh) if ($collate);
sub collate_and_clean {
my ($out_path, $file, $collate_fh) = @_;
my (@seqs, $seq);
parse_fasta("$out_path/$file/contigs.fa", \@seqs);
foreach $seq (@seqs) {
next if (length($seq->{'seq'}) < $min_len);
$seq->{'id'} = $file . "|" . $seq->{'id'};
print_fasta($collate_fh, $seq);
}
`rm $out_path/$file-g.output`;
`rm $out_path/$file-h.output`;
`rm -r $out_path/$file`;
}
sub parse_fasta {
my ($file, $seqs) = @_;
my ($fh, $line, $buf, $id, $seq);
open($fh, "<$file")
or die("Unable to open Velvet output file: $file, $!\n");
while ($line = <$fh>) {
chomp $line;
if (substr($line, 0, 1) eq ">") {
if (length($buf) > 0) {
$seq = {};
$seq->{'id'} = $id;
$seq->{'seq'} = $buf;
push(@{$seqs}, $seq);
$buf = "";
}
$id = substr($line, 1);
} else {
$buf .= $line;
}
}
if (length($buf) > 0 && length($id) > 0) {
$seq = {};
$seq->{'id'} = $id;
$seq->{'seq'} = $buf;
push(@{$seqs}, $seq);
}
close($fh);
}
sub print_fasta {
my ($fh, $seq) = @_;
my ($s);
print $fh ">", $seq->{'id'}, "\n";
$s = $seq->{'seq'};
while (length($s) > 60) {
print $fh substr($s, 0, 60), "\n";
$s = substr($s, 60);
}
print $fh $s, "\n" if (length($s) > 0);
}
sub build_file_list {
my ($files) = @_;
my (@ls, $line, $file, $path);
# Load a white list of files to process if it is supplied.
my @wl;
if (length($white_list) > 0) {
load_white_list(\@wl);
}
$path = length($paired_path) > 0 ? $paired_path : $single_path;
@ls = `ls -1 $path/`;
foreach $line (@ls) {
chomp $line;
next if (length($line) == 0);
next if ($line !~ /.+\.fa$/ && $line !~ /.+\.fasta$/);
($file) = ($line =~ /^(.+\.fas?t?a?)/);
if (scalar(@wl) > 0) {
next if (!grep(/^$file$/, @wl));
}
push(@{$files}, $file);
}
}
sub load_white_list {
my ($wl) = @_;
open(WHITE, "<" . $white_list)
or die("Unable to open white list file '$white_list': $!\n");
my $line = "";
while ($line = <WHITE>) {
chomp $line;
next if (length($line) == 0);
push(@{$wl}, $line);
}
close(WHITE);
}
sub parse_command_line {
while (@ARGV) {
$_ = shift @ARGV;
if ($_ =~ /^-s$/) { $single_path = shift @ARGV; }
elsif ($_ =~ /^-p$/) { $paired_path = shift @ARGV; }
elsif ($_ =~ /^-l$/) { $sanger_path = shift @ARGV; }
elsif ($_ =~ /^-o$/) { $out_path = shift @ARGV; }
elsif ($_ =~ /^-c$/) { $collate++; }
elsif ($_ =~ /^-W$/) { $white_list = shift @ARGV; }
elsif ($_ =~ /^-I$/) { $insert_len = shift @ARGV; }
elsif ($_ =~ /^-C$/) { $exp_cov = shift @ARGV; }
elsif ($_ =~ /^-T$/) { $cov_cut = shift @ARGV; }
elsif ($_ =~ /^-R$/) { $read_trk = shift @ARGV; }
elsif ($_ =~ /^-M$/) { $min_len = shift @ARGV; }
elsif ($_ =~ /^-H$/) { $hash_len = shift @ARGV; }
elsif ($_ =~ /^-P$/) { $paired++; }
elsif ($_ =~ /^-L$/) { $clean = 0; }
elsif ($_ =~ /^-e$/) { $exe_path = shift @ARGV; }
elsif ($_ =~ /^-v$/) { version(); exit(); }
elsif ($_ =~ /^-d$/) { $debug++; }
elsif ($_ =~ /^-h$/) { usage(); }
else {
print STDERR "Unknown command line option '$_'\n";
usage();
}
}
$single_path = substr($single_path, 0, -1) if (substr($single_path, -1) eq "/");
$paired_path = substr($paired_path, 0, -1) if (substr($paired_path, -1) eq "/");
$out_path = substr($out_path, 0, -1) if (substr($out_path, -1) eq "/");
$exe_path = substr($exe_path, 0, -1) if (substr($exe_path, -1) eq "/");
}
sub version {
print STDERR "exec_velvet.pl ", stacks_version, "\n";
}
sub usage {
version();
print STDERR <<EOQ;
exec_velvet.pl -p path -s path [-l path] -o path [-c] [-H len] [-P] [-I len] [-C exp_cov] [-T cov_cut]
[-W file_white_list] [-w marker_white_list] [-L] [-e path] [-d] [-h]
p: path to the paired-end FASTA files to assemble.
s: path to the single-end FASTA files to assemble.
l: path to long, sanger-style reads to assemble.
o: path to output the assembled files.
c: collate the resulting velvet runs into a single FASTA file, clean velvet directories.
W: a white list of files to process in the input path.
H: length of overlap required for reads (hash length, default 27).
P: process paired-end reads.
I: insert length (for paired-end reads; see velvet documentation).
C: expected coverage (for paired-end reads).
T: coverage cutoff.
M: minimum contig length, discard contigs shorter than this value.
R: turn on velvet's read tracking (uses additional memory).
L: leave velvet's intermediate files behind.
e: executable path, location of velvet programs.
h: display this help message.
d: turn on debug output.
EOQ
exit(0);
}
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