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#!/usr/bin/env perl
#
# Copyright 2010-2020, Julian Catchen <jcatchen@illinois.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 POSIX;
use File::Temp qw/ mktemp /;
use File::Spec;
use constant stacks_version => "_VERSION_";
use constant true => 1;
use constant false => 0;
my $dry_run = false;
my $exe_path = "_BINDIR_";
my $out_path = "";
my $popmap_path = "";
my $sample_path = "";
my $db = "";
my $min_cov = 0;
my $sample_id = 1;
my $gzip = false;
my $paired = false;
my $resume = false;
my $force_reexe = false;
my $genetic_map = false;
my $time = "";
my @parents;
my @progeny;
my @samples;
my (@_ustacks, @_cstacks, @_sstacks, @_tsv2bam, @_gstacks, @_populations);
my $cmd_str = $0 . " " . join(" ", @ARGV);
parse_command_line();
#
# Check for the existence of the necessary pipeline programs
#
foreach my $prog ("ustacks", "cstacks", "sstacks", "tsv2bam", "gstacks", "populations") {
die "Unable to find '" . $exe_path . $prog . "'.\n" if (!-e $exe_path . $prog);
}
my ($log, $log_fh, $sample);
my (@sample_list, %pop_ids, %pops, %grp_ids, %grps, %sample_ids);
parse_population_map(\@sample_list, \%pop_ids, \%pops, \%grp_ids, \%grps, \$genetic_map);
initialize_samples(\@parents, \@progeny, \@samples, \@sample_list, \%pop_ids, \%grp_ids);
#
# Open the log file
#
$log = "$out_path/denovo_map.log";
#
# If we are resuming, check if the log exists and append to it.
#
if ($resume == true) {
open($log_fh, ">>$log") or die("Unable to open log file '$log'; $!\n");
print $log_fh
"\n",
"denovo_map.pl version ", stacks_version, " resuming at ", strftime("%Y-%m-%d %H:%M:%S", (localtime(time))), "\n",
$cmd_str, "\n";
} else {
open($log_fh, ">$log") or die("Unable to open log file '$log'; $!\n");
print $log_fh
"denovo_map.pl version ", stacks_version, " started at ", strftime("%Y-%m-%d %H:%M:%S", (localtime(time))), "\n",
$cmd_str, "\n";
}
execute_stacks($log_fh, $sample_id, \@parents, \@progeny, \@samples, \%sample_ids);
print $log_fh "\ndenovo_map.pl completed at ", strftime("%Y-%m-%d %H:%M:%S", (localtime(time))), "\n";
close($log_fh);
sub check_return_value {
# $? is a 16 bit int. Exit code is given by `$? & 255` if the process was
# terminated by a signal, and by `$? >> 8` if it exited normally.
my ($rv, $log_fh) = @_;
if ($rv != 0) {
my $code = ($rv >> 8) & 127;
if ($rv & 255 || ($rv >> 8) > 127) {
$code += 128;
}
my $msg = "\ndenovo_map.pl: Aborted because the last command failed ($code";
if ($code == 129 || $code == 130 || $code == 131) {
$msg .= "/interrupted";
} elsif ($code == 137 || $code == 143) {
$msg .= "/killed";
} elsif ($code == 134) {
$msg .= "/SIGABRT";
} elsif ($code == 139) {
$msg .= "/segmentation fault";
}
$msg .= ")";
print $log_fh ($msg . ".\n");
print STDERR ($msg . "; see log file.\n");
exit 1;
}
}
sub execute_stacks {
my ($log_fh, $sample_id, $parents, $progeny, $samples, $sample_ids) = @_;
my (@results, @depths_of_cov, @prev_completed_ustacks);
my ($pop_cnt, $sample, $num_files, $i, $cmd, $pipe_fh);
my $minc = $min_cov > 0 ? " -m $min_cov" : "";
$i = 1;
$num_files = scalar(@{$parents}) + scalar(@{$progeny}) + scalar(@{$samples});
#
# Assemble RAD loci in each individual.
#
print STDERR "Indentifying unique stacks...\n";
print $log_fh "\nustacks\n==========\n";
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
print $log_fh "\nSample $i of $num_files '$sample->{'file'}'\n----------\n";
if (scalar(keys %{$sample_ids}) > 0) {
$sample_id = $sample_ids->{$sample->{'file'}};
}
#
# If resuming the pipeline, test if this sample already has been processed.
#
if ($resume == true and ustacks_sample_exists($sample, $out_path) == true) {
push(@prev_completed_ustacks, $sample);
print STDERR " Sample '$sample->{'file'}' already exists, will use existing data.\n";
print $log_fh "Sample '$sample->{'file'}' already exists, will use existing data.\n";
$i++;
$sample_id++;
next;
}
$cmd = $exe_path . "ustacks -t $sample->{'fmt'} -f $sample->{'path'} -o $out_path -i $sample_id" . $minc;
if ($sample->{'path'} !~ /$sample->{'file'}.$sample->{'suffix'}$/) {
# Guessing the sample name from the input path won't work.
$cmd .= " --name " . $sample->{'file'};
}
foreach (@_ustacks) {
$cmd .= " " . $_;
}
print STDERR " $cmd\n";
print $log_fh "$cmd\n";
if ($dry_run == false) {
open($pipe_fh, "$time $cmd 2>&1 |");
@results = ();
while (<$pipe_fh>) {
print $log_fh $_;
push(@results, $_);
}
close($pipe_fh);
check_return_value($?, $log_fh);
#
# Pull the depth of coverage from ustacks.
#
my $depthline = (grep(/^Final coverage/, @results))[0];
my ($depth) = ($depthline =~ /mean=([^;]+)/);
push(@depths_of_cov, [$sample->{'file'}, $depth]);
}
$i++;
$sample_id++;
}
write_depths_of_cov(\@depths_of_cov, $log_fh);
print STDERR "\n";
#
# Generate catalog of RAD loci.
#
print STDERR "Generating catalog...\n";
print $log_fh "\ncstacks\n==========\n";
#
# If a genetic map was specified (based on the population map), only place
# samples that are 'parent' into the catalog.
#
if ($genetic_map == true) {
my @params = ();
while (@_cstacks) {
my $param = pop(@_cstacks);
next if (substr($param, 0, 2) eq "-M");
push(@params, $param);
}
push(@_cstacks, @params);
foreach $sample (@{$parents}) {
push(@_cstacks, "-s " . $out_path . "/" . $sample->{'file'});
}
push(@_cstacks, "-o $out_path");
} else {
push(@_cstacks, "-P $out_path");
}
#
# If the pipeline is in --resume mode, check if all ustacks samples existed previously and
# if the catalog already exists.
#
if ($resume == true and catalog_exists(\@prev_completed_ustacks, $parents, $progeny, $samples, $out_path) == true) {
print STDERR " Catalog already exists, will use existing data.\n\n";
print $log_fh "Catalog already exists, will use existing data.\n\n";
} else {
#
# If --resume is enabled and we ended up re-executing cstacks, force everything downstream to re-execute.
#
$force_reexe = true if ($resume == true);
$cmd = $exe_path . "cstacks " . join(" ", @_cstacks);
print STDERR " $cmd\n\n";
print $log_fh "$cmd\n\n";
if ($dry_run == false) {
open($pipe_fh, "$time $cmd 2>&1 |");
while (<$pipe_fh>) {
print $log_fh $_;
if ($_ =~ /failed/i) { print STDERR "Catalog construction failed.\n"; exit(1); }
}
close($pipe_fh);
check_return_value($?, $log_fh);
}
}
#
# Match parents, progeny, or samples to the catalog.
#
print STDERR "Matching samples to the catalog...\n";
print $log_fh "\nsstacks\n==========\n";
if ($resume == true and sstacks_samples_exist($parents, $progeny, $samples, $out_path) == true) {
print STDERR " sstacks matches already exist, will use existing data.\n\n";
print $log_fh "sstacks matches already exist, will use existing data.\n\n";
} else {
#
# If --resume is enabled and we ended up re-executing sstacks, force everything downstream to re-execute.
#
$force_reexe = true if ($resume == true);
$cmd = $exe_path . "sstacks -P $out_path " . join(" ", @_sstacks);
print STDERR " $cmd\n\n";
print $log_fh "$cmd\n\n";
if ($dry_run == false) {
open($pipe_fh, "$time $cmd 2>&1 |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
check_return_value($?, $log_fh);
}
}
#
# Sort the reads according by catalog locus / run tsv2bam.
#
print STDERR "Sorting reads by RAD locus...\n";
print $log_fh "\ntsv2bam\n==========\n";
if ($resume == true and tsv2bam_samples_exist($parents, $progeny, $samples, $out_path) == true) {
print STDERR " tsv2bam samples already exist, will use existing data.\n\n";
print $log_fh "tsv2bam samples already exist, will use existing data.\n\n";
} else {
#
# If --resume is enabled and we ended up re-executing tsv2bam, force everything downstream to re-execute.
#
$force_reexe = true if ($resume == true);
my $file_paths = "";
if (length($popmap_path) == 0) {
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
$file_paths .= " -s $sample->{'file'}";
}
}
$cmd = $exe_path . "tsv2bam -P $out_path $file_paths " . join(" ", @_tsv2bam);
print STDERR " $cmd\n\n";
print $log_fh "$cmd\n\n";
if (!$dry_run) {
open($pipe_fh, "$time $cmd 2>&1 |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
check_return_value($?, $log_fh);
}
}
#
# Call genotypes / run gstacks.
#
print STDERR "Calling variants, genotypes and haplotypes...\n";
print $log_fh "\ngstacks\n==========\n";
if ($resume == true and gstacks_output_exists($out_path) == true) {
print STDERR " gstacks catalog already exists, will use existing data.\n\n";
print $log_fh "gstacks catalog already exists, will use existing data.\n\n";
} else {
$cmd = $exe_path . "gstacks -P $out_path " . join(" ", @_gstacks);
print STDERR " $cmd\n\n";
print $log_fh "$cmd\n\n";
if (!$dry_run) {
open($pipe_fh, "$time $cmd 2>&1 |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
check_return_value($?, $log_fh);
}
}
printf(STDERR "Calculating population-level summary statistics\n");
print $log_fh "\npopulations\n==========\n";
$cmd = $exe_path . "populations" . " -P $out_path " . join(" ", @_populations);
print STDERR " $cmd\n\n";
print $log_fh "$cmd\n\n";
if ($dry_run == 0) {
open($pipe_fh, "$time $cmd 2>&1 |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
check_return_value($?, $log_fh);
}
print STDERR "denovo_map.pl is done.\n";
print $log_fh "denovo_map.pl is done.\n";
}
sub ustacks_sample_exists {
my ($sample, $out_path) = @_;
my $path = $out_path . "/" . $sample->{'file'} . ".alleles.tsv.gz";
# Check the file exists and is not empty.
return false if (!-e $path || -z _);
my $fh;
open($fh, "gunzip -c $path | tail -n 1 |") or die("Unable to read ustacks file, '$path'\n");
my $line = <$fh>;
close($fh);
chomp $line;
return true if ($line =~ /^# ustacks completed/);
return false;
}
sub catalog_exists {
my ($prev_completed_ustacks, $parents, $progeny, $samples, $out_path) = @_;
return false if ($force_reexe == true);
my $sample_cnt = 0;
if ($genetic_map == true) {
$sample_cnt = scalar(@{$parents}) + scalar(@{$progeny});
} else {
$sample_cnt = scalar(@{$samples});
}
#
# If all ustacks samples have been processed, check if the catalog was completed.
#
if ($sample_cnt == scalar(@{$prev_completed_ustacks})) {
my $path = $out_path . "/catalog.tags.tsv.gz";
# Check the file exists and is not empty.
return false if (!-e $path || -z _);
my $path = $out_path . "/catalog.snps.tsv.gz";
return false if (!-e $path || -z _);
my $path = $out_path . "/catalog.alleles.tsv.gz";
return false if (!-e $path || -z _);
my $fh;
open($fh, "gunzip -c $path | tail -n 1 |") or die("Unable to read catalog file, '$path'\n");
my $line = <$fh>;
close($fh);
chomp $line;
return true if ($line =~ /^# cstacks completed/);
}
return false;
}
sub sstacks_samples_exist {
my ($parents, $progeny, $samples, $out_path) = @_;
return false if ($force_reexe == true);
my $sample_cnt = scalar(@{$parents}) + scalar(@{$progeny}) + scalar(@{$samples});
my $completed_cnt = 0;
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
my $path = $out_path . "/" . $sample->{'file'} . ".matches.tsv.gz";
# Check the file exists and is not empty.
return false if (!-e $path || -z _);
my $fh;
open($fh, "gunzip -c $path | tail -n 1 |") or die("Unable to read sstacks file, '$path'\n");
my $line = <$fh>;
close($fh);
chomp $line;
$completed_cnt++ if ($line =~ /^# sstacks completed/);
}
if ($sample_cnt == $completed_cnt) {
return true;
}
return false;
}
sub tsv2bam_samples_exist {
my ($parents, $progeny, $samples, $out_path) = @_;
return false if ($force_reexe == true);
my $sample_cnt = scalar(@{$parents}) + scalar(@{$progeny}) + scalar(@{$samples});
my $completed_cnt = 0;
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
my $path = $out_path . "/" . $sample->{'file'} . ".matches.bam";
# Check the file exists and is not empty.
$completed_cnt++ if (-e $path && !-z _);
}
if ($sample_cnt == $completed_cnt) {
my $path = $out_path . "/tsv2bam.log";
# Check the file exists and is not empty.
return false if (!-e $path || -z _);
my $fh;
open($fh, "tail -n 1 $path |") or die("Unable to read tsv2bam logfile, '$path'\n");
my $line = <$fh>;
close($fh);
chomp $line;
return true if ($line =~ /^tsv2bam is done./);
}
return false;
}
sub gstacks_output_exists {
my ($out_path) = @_;
return false if ($force_reexe == true);
my $path = $out_path . "/catalog.fa.gz";
# Check the file exists and is not empty.
return false if (!-e $path || -z _);
$path = $out_path . "/catalog.calls";
return false if (!-e $path || -z _);
$path = $out_path . "/gstacks.log";
return false if (!-e $path || -z _);
my $fh;
open($fh, "tail -n 1 $path |") or die("Unable to read tsv2bam logfile, '$path'\n");
my $line = <$fh>;
close($fh);
chomp $line;
return true if ($line =~ /^gstacks is done./);
return false;
}
sub parse_population_map {
my ($sample_list, $pop_ids, $pops, $grp_ids, $grps, $map) = @_;
my ($fh, @parts, $line, $sample);
return if (length($popmap_path) == 0);
open($fh, "<$popmap_path") or die("Unable to open population map, '$popmap_path', $!\n");
while ($line = <$fh>) {
chomp $line;
next if ($line =~ /^\s*#/);
@parts = split(/\t/, $line);
if (scalar(@parts) != 2 and scalar(@parts) != 3) {
die("Unable to parse population map, '$popmap_path' (expected 2 or 3 columns, found " . scalar(@parts) . "); at line:\n$line\n");
}
foreach my $part (@parts) {
$part =~ s/^\s*|\s*$//g;
}
push(@{$sample_list}, $parts[0]);
$pop_ids->{$parts[0]} = $parts[1];
$pops->{$parts[1]}++;
if (scalar(@parts) > 2) {
$grp_ids->{$parts[0]} = $parts[2];
$grps->{$parts[2]}++;
}
}
if (scalar(keys %{$grps}) == 0) {
$grps->{"1"}++;
foreach $sample (@{$sample_list}) {
$grp_ids->{$sample} = "1";
}
}
print STDERR "Parsed population map: ", scalar(@{$sample_list}), " files in ", scalar(keys %{$pops});
scalar(keys %{$pops}) == 1 ? print STDERR " population" : print STDERR " populations";
print STDERR " and ", scalar(keys %{$grps});
scalar(keys %{$grps}) == 1 ? print STDERR " group.\n" : print STDERR " groups.\n";
#
# Test for a genetic map.
#
my @k = keys %{$pops};
if (scalar(@k) == 2) {
if ( ($k[0] eq "parent" && $k[1] eq "progeny") ||
($k[0] eq "progeny" && $k[1] eq "parent") ) {
print STDERR "Genetic map detected from population map, only 'parent' samples will be loaded into the catalog.\n";
$$map = true;
}
}
close($fh);
}
sub initialize_samples {
my ($parents, $progeny, $samples, $sample_list, $pop_ids, $grp_ids) = @_;
if (scalar(@{$sample_list}) > 0 && scalar(@{$samples}) == 0) {
my @suffixes = ("fq", "fastq", "fq.gz", "fastq.gz", "fa", "fasta", "fa.gz", "fasta.gz");
my @fmts = ("fastq", "fastq", "gzfastq", "gzfastq", "fasta", "fasta", "gzfasta", "gzfasta");
#
# Read the samples in from the population map.
#
my ($i, $extension, $extension_pe);
my $first = true;
foreach $sample (@{$sample_list}) {
if ($first) {
$first = false;
my $found = false;
for ($i = 0; $i < scalar(@suffixes); $i++) {
if (!$paired && -e $sample_path . $sample . "." . $suffixes[$i]) {
$found = true;
$extension = "." . $suffixes[$i];
last;
} elsif (!$paired && -e $sample_path . $sample . ".1." . $suffixes[$i]) {
$found = true;
$extension = ".1." . $suffixes[$i];
last;
} elsif (-e $sample_path . $sample . ".1." . $suffixes[$i]) {
$found = true;
$extension = ".1." . $suffixes[$i];
$extension_pe = ".2." . $suffixes[$i];
last;
}
}
if (!$found) {
print STDERR "Error: Failed to find the first reads file '$sample_path$sample(.1).(fq|fastq|fa|fasta)(.gz)'.\n";
exit 1;
}
if ($i == 2 || $i == 3 || $i == 6 || $i == 7) {
$gzip = true;
}
}
my ($path, $path_pe);
$path = $sample_path . $sample . $extension;
$path_pe = ($paired ? $sample_path . $sample . $extension_pe : "");
die("Error: Failed to open single-end file '$path'.\n") if (! -e $path);
die("Error: Failed to open paired-end file '$path_pe'.\n") if ($paired && ! -e $path_pe);
die("Unable to find an entry for '" . $sample . "' in the population map, '$popmap_path'.\n") if (!defined($pop_ids->{$sample}));
if ($genetic_map == true) {
if ($pop_ids->{$sample} eq "parent") {
push(@{$parents}, {'path' => $path,
'file' => $sample,
'suffix' => $suffixes[$i],
'type' => "parent",
'fmt' => $fmts[$i],
'path_pe' => $path_pe});
} else {
push(@{$progeny}, {'path' => $path,
'file' => $sample,
'suffix' => $suffixes[$i],
'type' => "progeny",
'fmt' => $fmts[$i],
'path_pe' => $path_pe});
}
} else {
push(@{$samples}, {'path' => $path,
'file' => $sample,
'suffix' => $suffixes[$i],
'type' => "sample",
'fmt' => $fmts[$i],
'path_pe' => $path_pe});
}
}
}
#
# If a population map was specified, make sure all samples in the list were found (and vice versa) and assign popualtion IDs.
#
my %sample_hash;
foreach $sample (@{$samples}, @{$parents}, @{$progeny}) {
$sample_hash{$sample->{'file'}}++;
if (!defined($pop_ids->{$sample->{'file'}})) {
die("Unable to find an entry for '" . $sample->{'file'} . "' in the population map, '$popmap_path'.\n");
} else {
$sample->{'pop_id'} = $pop_ids->{$sample->{'file'}};
}
if (!defined($grp_ids->{$sample->{'file'}})) {
die("Unable to find an entry for '" . $sample->{'file'} . "' in the population map, '$popmap_path'.\n");
} else {
$sample->{'grp_id'} = $grp_ids->{$sample->{'file'}};
}
}
foreach $sample (@{$sample_list}) {
if (!defined($sample_hash{$sample})) {
die("Unable to find a file corresponding to the population map entry '" . $sample . "' in the population map, '$popmap_path'.\n");
}
}
#
# Check that no duplicate files were specified.
#
my (%files, $file);
foreach $file (@{$parents}, @{$progeny}, @{$samples}) {
$files{$file}++;
}
foreach $file (keys %files) {
if ($files{$file} > 1) {
die("A duplicate file was specified which may create undefined results, '$file'\n");
}
}
if (scalar(@{$samples}) > 0) {
print STDERR "Found ", scalar(@{$samples}), " sample file(s).\n\n";
} else {
print STDERR
"Found ", scalar(@{$parents}), " parental file(s).\n",
"Found ", scalar(@{$progeny}), " progeny file(s).\n\n";
}
if ( scalar(@{$samples}) > 0 && (scalar(@{$parents}) > 0 || scalar(@{$progeny}) > 0) ) {
die("Both samples and parents/progeny were specified either on the command line (-s/-r/-p) or within the population map. Only one of the other may be specified.\n");
}
}
sub write_results {
my ($results, $log_fh) = @_;
my $line;
foreach $line (@{$results}) {
if ($line =~ /\r/) {
$line =~ s/^.+\r(.*\n)$/\1/;
}
print $log_fh $line;
}
}
sub write_depths_of_cov {
my ($depths, $log_fh) = @_;
print STDERR "\nDepths of Coverage for Processed Samples:\n";
print $log_fh "\nDepths of Coverage for Processed Samples:\n";
foreach $a (@{$depths}) {
print STDERR $a->[0], ": ", $a->[1], "x\n";
print $log_fh $a->[0], ": ", $a->[1], "x\n";
}
}
sub parse_command_line {
my ($arg);
my $ustacks_mismatch = -1;
my $cstacks_mismatch = -1;
while (@ARGV) {
$_ = shift @ARGV;
if ($_ =~ /^-v$/ || $_ =~ /^--version$/) { version(); exit 1; }
elsif ($_ =~ /^-h$/) { usage(); }
elsif ($_ =~ /^-d$/ || $_ =~ /^--dry-run$/) { $dry_run = true; }
elsif ($_ =~ /^-o$/ || $_ =~ /^--out-path$/) { $out_path = shift @ARGV; }
elsif ($_ =~ /^-e$/) { $exe_path = shift @ARGV; }
elsif ($_ =~ /^-m$/) { $min_cov = shift @ARGV; }
elsif ($_ =~ /^--resume$/) { $resume = true; }
elsif ($_ =~ /^--paired$/) { $paired = true; }
elsif ($_ =~ /^--samples$/) { $sample_path = shift @ARGV; }
elsif ($_ =~ /^-O$/ || $_ =~ /^--popmap$/) {
$popmap_path = shift @ARGV;
push(@_cstacks, "-M " . $popmap_path);
push(@_sstacks, "-M " . $popmap_path);
push(@_tsv2bam, "-M " . $popmap_path);
push(@_gstacks, "-M " . $popmap_path);
push(@_populations, "-M " . $popmap_path);
} elsif ($_ =~ /^-T$/ || $_ =~ /^--threads/) {
$arg = shift @ARGV;
push(@_ustacks, "-p " . $arg);
push(@_cstacks, "-p " . $arg);
push(@_sstacks, "-p " . $arg);
push(@_tsv2bam, "-t " . $arg);
push(@_gstacks, "-t " . $arg);
push(@_populations, "-t " . $arg);
} elsif ($_ =~ /^-M$/) {
$ustacks_mismatch = shift(@ARGV);
push(@_ustacks, "-M " . $ustacks_mismatch);
} elsif ($_ =~ /^-N$/) {
push(@_ustacks, "-N " . shift @ARGV);
} elsif ($_ =~ /^-n$/) {
$cstacks_mismatch = shift @ARGV;
push(@_cstacks, "-n " . $cstacks_mismatch);
} elsif ($_ =~ /^--rm-pcr-duplicates$/) {
push(@_gstacks, "--rm-pcr-duplicates");
} elsif ($_ =~ /^--var-alpha$/) {
push(@_gstacks, "--var-alpha " . shift @ARGV);
} elsif ($_ =~ /^--gt-alpha$/) {
push(@_gstacks, "--gt-alpha " . shift @ARGV);
} elsif ($_ =~ /^-r$/ || $_ =~ /^--min-samples-per-pop$/) {
push(@_populations, "--min-samples-per-pop " . shift @ARGV);
} elsif ($_ =~ /^-p$/ || $_ =~ /^--min-populations$/) {
push(@_populations, "--min-populations " . shift @ARGV);
} elsif ($_ =~ /^-X$/) {
#
# Pass an arbitrary command-line option to a pipeline program.
#
# Command line option must be of the form '-X "program:option"'
#
$arg = shift @ARGV;
my ($prog, $opt) = ($arg =~ /^(\w+):(.+)$/);
if ($prog eq "ustacks") {
push(@_ustacks, $opt);
} elsif ($prog eq "cstacks") {
push(@_cstacks, $opt);
} elsif ($prog eq "sstacks") {
push(@_sstacks, $opt);
} elsif ($prog eq "tsv2bam") {
push(@_tsv2bam, $opt);
} elsif ($prog eq "gstacks") {
push(@_gstacks, $opt);
} elsif ($prog eq "populations") {
push(@_populations, $opt);
} else {
print STDERR "Unknown pipeline program, '$arg'\n";
usage();
}
} elsif ($_ =~ /^--time-components$/) {
$time = '/usr/bin/time';
if (! -e $time) {
die "Error: '$time': No such file or directory.\n";
}
} else {
print STDERR "Unknown command line option: '$_'\n";
usage();
}
}
$exe_path = $exe_path . "/" if (substr($exe_path, -1) ne "/");
$out_path = substr($out_path, 0, -1) if (substr($out_path, -1) eq "/");
if (length($popmap_path) == 0) {
print STDERR "You must specify a population map that lists your sample names (--popmap).\n";
usage();
}
if (length($sample_path) == 0) {
print STDERR "You must specify the path to the directory containing the samples (--samples).\n";
usage();
}
if (length($sample_path) > 0) {
$sample_path .= "/" if (substr($sample_path, -1) ne "/");
}
if ($paired == true) {
push(@_tsv2bam, "-R $sample_path");
}
#
# By default, we want ustacks -M to equal cstacks -n.
#
if ($cstacks_mismatch == -1 && $ustacks_mismatch > 0) {
push(@_cstacks, "-n " . $ustacks_mismatch);
}
}
sub version {
print STDERR "denovo_map.pl ", stacks_version, "\n";
}
sub usage {
version();
print STDERR <<EOQ;
denovo_map.pl --samples dir --popmap path --out-path dir [--paired [--rm-pcr-duplicates]] (assembly options) (filtering options) [-X prog:"opts" ...]
Input/Output files:
--samples: path to the directory containing the samples reads files.
--popmap: path to a population map file (format is "<name> TAB <pop>", one sample per line).
-o,--out-path: path to an output directory.
General options:
-X: additional options for specific pipeline components, e.g. -X "populations: --min-maf 0.05".
-T, --threads: the number of threads/CPUs to use (default: 1).
--dry-run: dry run. Do not actually execute anything, just print the commands that would be executed.
--resume: resume executing the pipeline from a previous run.
Stack assembly options:
-M: number of mismatches allowed between stacks within individuals (for ustacks).
-n: number of mismatches allowed between stacks between individuals (for cstacks; default 1; suggested: set to ustacks -M).
SNP model options:
--var-alpha: significance level at which to call variant sites (for gstacks; default: 0.05).
--gt-alpha: significance level at which to call genotypes (for gstacks; default: 0.05).
Paired-end options:
--paired: after assembling RAD loci, assemble mini-contigs with paired-end reads.
--rm-pcr-duplicates: remove all but one set of read pairs of the same sample that have
the same insert length.
Population filtering options:
-r,--min-samples-per-pop: minimum percentage of individuals in a population required to process a locus for that population (for populations; default: 0)
-p,--min-populations: minimum number of populations a locus must be present in to process a locus (for populations; default: 1)
Miscellaneous:
--time-components (for benchmarking)
EOQ
exit 1;
}
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