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#!/usr/bin/env perl
#
# Copyright (C) 2013-2025 Genome Research Ltd.
#
# Author: Petr Danecek <pd3@sanger.ac.uk>
#
# 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 Carp;
use Cwd qw/ abs_path /;
use FindBin;
use lib "$FindBin::Bin";
use Getopt::Long;
use File::Temp;
use IO::Handle;
my $opts = parse_params();
test_reference($opts);
test_reference($opts, threads=>2);
test_bgzip($opts);
test_faidx($opts, threads=>2);
test_fqidx($opts, threads=>2);
test_dict($opts);
test_index($opts);
test_index($opts, threads=>2);
test_mpileup($opts);
test_usage($opts, cmd=>'samtools');
test_view($opts);
test_head($opts);
test_cat($opts);
test_import($opts);
test_bam2fq($opts);
test_bam2fq($opts, threads=>2);
test_depad($opts);
test_stats($opts);
test_merge($opts);
test_merge($opts, threads=>2);
test_sort($opts);
test_sort($opts, threads=>2);
test_collate($opts);
test_collate($opts, threads=>2);
test_fixmate($opts);
test_fixmate($opts, threads=>2);
test_calmd($opts);
test_calmd($opts, threads=>2);
test_idxstat($opts);
test_quickcheck($opts);
test_reheader($opts);
test_addrprg($opts);
test_addrprg($opts, threads=>2);
test_markdup($opts);
test_markdup($opts, threads=>2);
test_bedcov($opts);
test_split($opts);
test_split($opts, threads=>2);
test_large_positions($opts);
test_ampliconclip($opts);
test_ampliconclip($opts, threads=>2);
test_ampliconstats($opts, threads=>2);
test_reset($opts);
test_checksum($opts);
test_checksum($opts, threads=>2);
print "\nNumber of tests:\n";
printf " total .. %d\n", $$opts{nok}+$$opts{nfailed}+$$opts{nxfail}+$$opts{nxpass};
printf " passed .. %d\n", $$opts{nok};
printf " failed .. %d\n", $$opts{nfailed};
printf " expected failure .. %d\n", $$opts{nxfail};
printf " unexpected pass .. %d\n", $$opts{nxpass};
print "\n";
# Exit non-zero if there is a failure or an unexpected pass. In the case
# of an unexpected pass, the test script itself is at fault and should
# be updated to expect a pass instead of failure.
exit ($$opts{nfailed} > 0 || $$opts{nxpass} > 0);
#--------------------
sub error
{
my (@msg) = @_;
if ( scalar @msg ) { confess @msg; }
print
"About: samtools/htslib consistency test script\n",
"Usage: test.pl [OPTIONS]\n",
"Options:\n",
" -e, --exec <tool>=[<path>] Path to use for specified tool executable.\n",
" -r, --redo-outputs Recreate expected output files.\n",
" -t, --temp-dir <path> When given, temporary files will not be removed.\n",
" -h, -?, --help This help message.\n",
"\n";
exit 1;
}
sub tempfile {
my ($fh, $name) = File::Temp::tempfile(@_);
if (wantarray) {
if ($^O =~ /^(?:cygwin|msys|MSWin32)/) {
$name = abs_path($name);
}
return ($fh, $name);
}
return $fh;
}
sub cygpath {
my ($path) = @_;
$path = `cygpath -m $path`;
$path =~ s/\r?\n//;
return $path
}
sub tempdir
{
my $dir = File::Temp::tempdir(@_);
if ($^O =~ /^(cygwin|msys)/) {
$dir = cygpath($dir);
} elsif ($^O eq 'MSWin32') {
$dir =~ s/\\/\//g;
}
return $dir;
}
sub parse_params
{
my $opts = { bgzip=>"bgzip", keep_files=>0, nok=>0, nfailed=>0, nxfail => 0, nxpass => 0 };
my $help;
Getopt::Long::Configure('bundling');
my $ret = GetOptions (
'e|exec=s' => sub {
my ($tool, $path) = split /=/, $_[1];
if ($^O eq 'MSWin32' && $path !~ /\.exe$/) {
$path .= '.exe';
}
$$opts{$tool} = abs_path($path) if $path;
},
't|temp-dir:s' => \$$opts{keep_files},
'r|redo-outputs' => \$$opts{redo_outputs},
'h|?|help' => \$help
);
if ( !$ret or $help ) { error(); }
$$opts{tmp} = $$opts{keep_files} ? $$opts{keep_files} : tempdir(CLEANUP => 1);
$$opts{path} = $FindBin::RealBin;
$$opts{bin} = $FindBin::RealBin;
if ($^O =~ /^(cygwin|msys)/) {
$$opts{path} = cygpath($$opts{path});
$$opts{bin} = cygpath($$opts{bin});
}
$$opts{bin} =~ s{/test/?$}{};
if ( $$opts{keep_files} ) { cmd("mkdir -p $$opts{keep_files}"); }
else
{
$SIG{TERM} = $SIG{INT} = sub { clean_files($opts); };
}
$$opts{diff} = "diff" . ($^O =~ /^(?:cygwin|msys|MSWin32)/ ? " -b":"");
return $opts;
}
sub clean_files
{
my ($opts) = @_;
print STDERR "Signal caught, cleaning and exiting...\n";
`rm -rf $$opts{tmp}`;
}
sub _cmd
{
my ($cmd, $args) = @_;
my $kid_io;
my $out;
my $err;
my ($err_fh, $err_filename) = tempfile(UNLINK => 1);
my $pid = open($kid_io, "-|", 'bash', '-o','pipefail','-c', "($cmd) 2> $err_filename");
if ( !defined $pid ) { error("Cannot fork: $!"); }
if ($pid)
{
# parent
binmode($kid_io);
binmode($err_fh);
local $/;
$out = <$kid_io>;
close($kid_io);
my $child_retval = $?;
$err = <$err_fh>;
close ($err_fh);
$out =~ s/\x0d\x0a/\x0a/g if (!$args->{binary});
$err =~ s/\x0d\x0a/\x0a/g if (!$args->{binary_err});
$out =~ s/e([-+])0(\d\d)/e$1$2/g if ($args->{exp_fix});
return ($child_retval, $out, $err);
}
}
sub cmd
{
my ($cmd, $args) = @_;
my ($ret,$out,$err) = _cmd($cmd, $args);
if ( $ret ) { error("The command failed [$ret]: $cmd\n", "out:$out\n", "err:$err\n"); }
return $out;
}
# test harness for a command
# %args out=> expected output (must be present)
# err=> expected stderr output (optional)
# cmd=> command to test
# expect_fail=> as per passed()/failed()
# want_fail=> consider passed() if cmd() returns non-zero
# out_map => map output filenames to their expected result file (can be used alongside out)
# hskip => number of header lines to ignore during diff
# ignore_pg_header => remove @PG header lines
# reorder_header => ignore inconsequential header line order changes
sub test_cmd
{
my ($opts,%args) = @_;
if ( !exists($args{out}) )
{
if ( !exists($args{in}) ) { error("FIXME: expected out or in key\n"); }
$args{out} = "$args{in}.out";
}
my ($package, $filename, $line, $test)=caller(1);
$test =~ s/^.+:://;
print "$test:\n";
print "\t$args{cmd}\n";
my ($ret,$out,$err) = _cmd("$args{cmd}", \%args);
if ( $args{want_fail}? ($ret == 0) : ($ret != 0) ) { failed($opts,%args,msg=>$test,reason=>"ERR: $err"); return; }
if ( $$opts{redo_outputs} && -e "$$opts{path}/$args{out}" )
{
rename("$$opts{path}/$args{out}","$$opts{path}/$args{out}.old");
open(my $fh,'>',"$$opts{path}/$args{out}") or error("$$opts{path}/$args{out}: $!");
binmode($fh);
print $fh $out;
close($fh);
my ($ret,$out) = _cmd("$$opts{diff} -q $$opts{path}/$args{out} $$opts{path}/$args{out}.old");
if ( !$ret && $out eq '' ) { unlink("$$opts{path}/$args{out}.old"); }
else
{
print "\tthe expected output changed, saving:\n";
print "\t old .. $$opts{path}/$args{out}.old\n";
print "\t new .. $$opts{path}/$args{out}\n";
}
}
if ( $$opts{redo_outputs} && -e "$$opts{path}/$args{err}" )
{
rename("$$opts{path}/$args{err}","$$opts{path}/$args{err}.old");
open(my $fh,'>',"$$opts{path}/$args{err}") or error("$$opts{path}/$args{err}: $!");
binmode($fh);
print $fh $err;
close($fh);
my ($ret,$out) = _cmd("$$opts{diff} -q $$opts{path}/$args{err} $$opts{path}/$args{err}.old");
if ( !$ret && $err eq '' ) { unlink("$$opts{path}/$args{err}.old"); }
else
{
print "\tthe expected stderr output changed, saving:\n";
print "\t old .. $$opts{path}/$args{err}.old\n";
print "\t new .. $$opts{path}/$args{err}\n";
}
}
# check stdout
my $exp = '';
if ( open(my $fh,'<',"$$opts{path}/$args{out}") )
{
binmode($fh);
local $/;
$exp = <$fh>;
close($fh);
}
elsif ( !$$opts{redo_outputs} ) { failed($opts,%args,msg=>$test,reason=>"$$opts{path}/$args{out}: $!"); return; }
if ($args{reorder_header}) {
$exp = reorder_headers($exp, $out);
}
if ($args{ignore_pg_header}) {
$out =~ s/(^|\n)(?:\@PG\t[^\n]*\n)+/$1/sg;
$exp =~ s/(^|\n)(?:\@PG\t[^\n]*\n)+/$1/sg;
}
if ( $exp ne $out )
{
open(my $fh,'>',"$$opts{path}/$args{out}.new") or error("$$opts{path}/$args{out}.new");
binmode($fh);
print $fh $out;
close($fh);
if ( !-e "$$opts{path}/$args{out}" )
{
rename("$$opts{path}/$args{out}.new","$$opts{path}/$args{out}") or error("rename $$opts{path}/$args{out}.new $$opts{path}/$args{out}: $!");
print "\tthe file with expected output does not exist, creating new one:\n";
print "\t\t$$opts{path}/$args{out}\n";
}
else
{
failed($opts,%args,msg=>$test,reason=>"The outputs stdout differ:\n\t\t$$opts{path}/$args{out}\n\t\t$$opts{path}/$args{out}.new");
# system("hexdump -C $$opts{path}/$args{out}");
# system("hexdump -C $$opts{path}/$args{out}.new");
}
return;
}
# check stderr
if ( exists($args{err}) ) {
my $exp_err = '';
if ( open(my $fh,'<',"$$opts{path}/$args{err}") )
{
binmode($fh);
my @exp = <$fh>;
$exp_err = join('',@exp);
close($fh);
}
elsif ( !$$opts{redo_outputs} ) { failed($opts,%args,msg=>$test,reason=>"$$opts{path}/$args{err}: $!"); return; }
if ( $exp_err ne $err )
{
open(my $fh,'>',"$$opts{path}/$args{err}.new") or error("$$opts{path}/$args{err}.new");
binmode($fh);
print $fh $err;
close($fh);
if ( !-e "$$opts{path}/$args{err}" )
{
rename("$$opts{path}/$args{err}.new","$$opts{path}/$args{err}") or error("rename $$opts{path}/$args{err}.new $$opts{path}/$args{err}: $!");
print "\tthe file with expected output does not exist, creating new one:\n";
print "\t\t$$opts{path}/$args{err}\n";
}
else
{
failed($opts,%args,msg=>$test,reason=>"The outputs stderr differ:\n\t\t$$opts{path}/$args{err}\n\t\t$$opts{path}/$args{err}.new");
}
return;
}
}
# check other output files
if( exists($args{out_map}) )
{
while ( my($out_actual, $out_expected) = each %{$args{out_map}} )
{
my $exp = '';
if ( open(my $fh,'<',"$$opts{path}/$out_expected") )
{
binmode($fh);
local $/;
$exp = <$fh>;
close($fh);
}
elsif ( !$$opts{redo_outputs} ) { failed($opts,%args,msg=>$test,reason=>"$$opts{path}/$out_expected: $!"); return; }
my $out = '';
if ( open(my $fh,'<',"$$opts{path}/$out_actual") )
{
binmode($fh);
if( exists($args{hskip}) ){
# Strip hskip lines to allow match to the expected output
for (my $i = 0; $i < $args{hskip}; $i++) {
my $ignore = <$fh>;
}
}
local $/;
$out = <$fh>;
$out =~ s/\x0d\x0a/\x0a/g if (!$args{binary});
$out =~ s/e\+0(\d\d)/e+$1/g if ($args{exp_fix});
close($fh);
}
elsif ( !$$opts{redo_outputs} ) { failed($opts,%args,msg=>$test,reason=>"$$opts{path}/$out_actual: $!"); return; }
if ($args{ignore_pg_header}) {
$out =~ s/(^|\n)\@PG\t[^\n]*\n/$1/mg;
$exp =~ s/(^|\n)\@PG\t[^\n]*\n/$1/mg;
}
if ($args{reorder_header}) {
$exp = reorder_headers($exp, $out);
}
if ( $exp ne $out )
{
open(my $fh,'>',"$$opts{path}/$out_expected.new") or error("$$opts{path}/$out_expected.new");
binmode($fh);
print $fh $out;
close($fh);
if ( !-e "$$opts{path}/$out_expected" )
{
rename("$$opts{path}/$out_expected.new","$$opts{path}/$out_expected") or error("rename $$opts{path}/$out_expected.new $$opts{path}/$out_expected: $!");
print "\tthe file with expected output does not exist, creating new one:\n";
print "\t\t$$opts{path}/$out_expected\n";
}
else
{
failed($opts,%args,msg=>$test,reason=>"The output files differ:\n\t\t$$opts{path}/$out_expected\n\t\t$$opts{path}/$out_expected.new");
}
return;
}
unlink("$$opts{path}/$out_actual");
}
}
passed($opts,%args,msg=>$test);
}
# Record the success or failure of a test. $opts is the global settings hash;
# %args is a list of key => value pairs that may include:
# msg => Message describing the test (currently unused)
# reason => Description of why the test failed (only for failed())
# expect_fail => If set, records failed as xfail and passed as xpass
sub failed
{
my ($opts, %args) = @_;
my $reason = exists $args{reason}? "\t$args{reason}\n" : "";
if (!$args{expect_fail}) {
$$opts{nfailed}++;
print "\n"; STDOUT->flush();
print STDERR "$reason.. failed ...\n"; STDERR->flush();
print "\n";
}
else { $$opts{nxfail}++; print "\n$reason.. expected failure\n\n"; }
}
sub passed
{
my ($opts, %args) = @_;
if (!$args{expect_fail}) { $$opts{nok}++; print ".. ok\n\n"; }
else {
$$opts{nxpass}++;
STDOUT->flush();
print STDERR ".. unexpected pass\n"; STDERR->flush();
print "\n";
}
}
sub is_file_newer
{
my ($afile,$bfile) = @_;
my (@astat) = stat($afile) or return 0;
my (@bstat) = stat($bfile) or return 0;
if ( $astat[9]>$bstat[9] ) { return 1 }
return 0;
}
sub reorder_headers
{
my ($exp, $out) = @_;
my %headers;
while ($exp =~ /^\@(..)(.*)\n/mg) {
my ($type, $vals) = ($1, $2);
if ($type eq 'PG' || $type eq 'RG') {
my ($id) = $vals =~ /\tID:([^\t]+)/;
push(@{$headers{$type}->{$id || ""}}, $vals);
} else {
push(@{$headers{$type}}, $vals || "");
}
}
$exp =~ s/^\@...*\n//mg;
my $reorder = "";
while ($out =~ /^\@(..)(.*)\n/mg) {
my ($type, $vals) = ($1, $2);
if ($type eq 'PG' || $type eq 'RG') {
my ($id) = $vals =~ /\tID:([^\t]+)/;
if (exists($headers{$type}->{$id || ""})
&& @{$headers{$type}->{$id}}) {
my $v = shift(@{$headers{$type}->{$id}});
$reorder .= "\@$type$v\n";
}
} elsif (exists($headers{$type}) && @{$headers{$type}}) {
my $v = shift(@{$headers{$type}});
$reorder .= "\@$type$v\n";
}
}
# Deal with any left-overs
foreach my $type (sort keys %headers) {
if ($type eq 'PG' || $type eq 'RG') {
foreach my $id (sort keys %{$headers{$type}}) {
foreach my $v (@{$headers{$type}->{$id}}) {
$reorder .= "\@$type$v\n";
}
}
} else {
foreach my $v (@{$headers{$type}}) {
$reorder .= "\@$type$v\n";
}
}
}
return $reorder . $exp;
}
# The tests --------------------------
sub test_bgzip
{
my ($opts,%args) = @_;
# Create test data: The beginning of each line gives the 0-based offset (including '\n's)
#
open(my $fh,'>',"$$opts{tmp}/bgzip.dat") or error("$$opts{tmp}/bgzip.dat: $!");
binmode($fh);
my $ntot = 1_000_000;
my $nwr = 0;
while ($nwr < $ntot)
{
my $out = sprintf("%d\n", $nwr);
$nwr += length($out);
print $fh $out;
}
close($fh);
cmd("cat $$opts{tmp}/bgzip.dat | $$opts{bgzip} -ci -I$$opts{tmp}/bgzip.dat.gz.gzi > $$opts{tmp}/bgzip.dat.gz");
# Run tests
my ($test,$out);
$test = "$$opts{bgzip} -c -b 65272 -s 5 $$opts{tmp}/bgzip.dat.gz";
print "$test\n";
$out = cmd($test);
if ( $out ne '65272' ) { failed($opts,msg=>$test,reason=>"Expected \"65272\" got \"$out\"\n"); }
else { passed($opts,msg=>$test); }
$test = "$$opts{bgzip} -c -b 979200 -s 6 $$opts{tmp}/bgzip.dat.gz";
print "$test\n";
$out = cmd($test);
if ( $out ne '979200' ) { failed($opts,msg=>$test,reason=>"Expected \"979200\" got \"$out\"\n"); }
else { passed($opts,msg=>$test); }
$test = "$$opts{bgzip} -c -b 652804 -s 6 $$opts{tmp}/bgzip.dat.gz";
print "$test\n";
$out = cmd($test);
if ( $out ne '652804' ) { failed($opts,msg=>$test,reason=>"Expected \"652804\" got \"$out\"\n"); }
else { passed($opts,msg=>$test); }
}
# For faidx testing: code numbers as base 4 ACGT strings
sub faidx_num_to_seq
{
my ($dec) = @_;
my $out = '';
my @base = qw(A C G T);
while ( $dec>=0 )
{
my $r = $dec % 4;
$out = $base[$r] . $out;
$dec = int( ($dec - $r) / 4 );
if ( !$dec ) { last; }
}
return $out;
}
sub faidx_seq_to_num
{
my ($seq) = @_;
my $out = 0;
my $len = length($seq);
my %base = ( A=>0, C=>1, G=>2, T=>3 );
for (my $i=0; $i<$len; $i++)
{
my $b = substr($seq,$i,1);
$out += $base{$b} * 4**($len-$i-1);
}
return $out;
}
sub faidx_wrap
{
my ($seq,$width) = @_;
my $out;
while ( length($seq) )
{
$out .= substr($seq,0,60,'');
$out .= "\n";
}
return $out;
}
sub test_faidx
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
# Create test data: The fake sequence consists of sequence offsets coded
# into A,C,G,T and separated with Ns. The offsets are 1-based.
#
open(my $fh,'>',"$$opts{tmp}/faidx.fa") or error("$$opts{tmp}/faidx.fa: $!");
my $ntot = 100_000;
my @dat = qw(A C G T);
for (my $seq=1; $seq<=3; $seq++)
{
my $nwr = 1;
my $out = '';
while ($nwr < $ntot)
{
my $tmp = faidx_num_to_seq($nwr) . 'N';
$out .= $tmp;
$nwr += length($tmp);
}
print $fh ">$seq\n";
print $fh faidx_wrap($out);
}
close($fh);
# Run tests: index and retrieval from plain text and compressed files
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa");
cmd("cat $$opts{tmp}/faidx.fa | $$opts{bgzip} -ci -I $$opts{tmp}/faidx.fa.gz.gzi > $$opts{tmp}/faidx.fa.gz");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa.gz");
# Write to file
cmd("$$opts{bin}/samtools faidx --length 5 $$opts{tmp}/faidx.fa 1:1-104 > $$opts{tmp}/output_faidx_base.fa");
cmd("$$opts{bin}/samtools faidx --length 5 $$opts{tmp}/faidx.fa 1 > $$opts{tmp}/output_faidx_base.1.fa");
cmd("$$opts{bin}/samtools faidx --length 5 --output $$opts{tmp}/output_faidx.fa $$opts{tmp}/faidx.fa 1:1-104 && $$opts{diff} $$opts{tmp}/output_faidx.fa $$opts{tmp}/output_faidx_base.fa");
# Write to bgzip file
cmd("$$opts{bin}/samtools faidx --length 5 --output $$opts{tmp}/output_faidx.fa.1.gz $$opts{tmp}/faidx.fa 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_faidx.fa.1.gz && $$opts{diff} $$opts{tmp}/output_faidx.fa.1 $$opts{tmp}/output_faidx_base.fa");
cmd("$$opts{bin}/samtools faidx --length 5 --output $$opts{tmp}/output_faidx.fa.2.bgz $$opts{tmp}/faidx.fa 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_faidx.fa.2.bgz && $$opts{diff} $$opts{tmp}/output_faidx.fa.2 $$opts{tmp}/output_faidx_base.fa");
cmd("$$opts{bin}/samtools faidx --length 5 --output $$opts{tmp}/output_faidx.fa.3.bgzf $$opts{tmp}/faidx.fa 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_faidx.fa.3.bgzf && $$opts{diff} $$opts{tmp}/output_faidx.fa.3 $$opts{tmp}/output_faidx_base.fa");
# Write to bgzip file with compression level
cmd("$$opts{bin}/samtools faidx --length 5 --output $$opts{tmp}/output_faidx.fa.4.gz $$opts{tmp}/faidx.fa 1 --output-fmt-opt=\"level=4\" && $$opts{bgzip} -df $$opts{tmp}/output_faidx.fa.4.gz && $$opts{diff} $$opts{tmp}/output_faidx.fa.4 $$opts{tmp}/output_faidx_base.1.fa");
# Write to bgzip file with thread
cmd("$$opts{bin}/samtools faidx ${threads} --length 5 --output $$opts{tmp}/output_faidx.fa.5.gz $$opts{tmp}/faidx.fa 1 && $$opts{bgzip} -df $$opts{tmp}/output_faidx.fa.5.gz && $$opts{diff} $$opts{tmp}/output_faidx.fa.5 $$opts{tmp}/output_faidx_base.1.fa");
# Write indices to a file
cmd("$$opts{bin}/samtools faidx --fai-idx $$opts{tmp}/fa_test.fai --gzi-idx $$opts{tmp}/fa_test.gzi $$opts{tmp}/faidx.fa.gz");
cmd("$$opts{diff} $$opts{tmp}/faidx.fa.fai $$opts{tmp}/fa_test.fai");
cmd("$$opts{diff} $$opts{tmp}/faidx.fa.gz.gzi $$opts{tmp}/fa_test.gzi");
#with write-index
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa 1 --write-index"); #ignores write-index
cmd("echo \"1\n2\n3\" > $$opts{tmp}/1.reg"); #create region file
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa --write-index -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fa"); #writes and indexes
cmd("$$opts{diff} $$opts{tmp}/faidx.fa.fai $$opts{tmp}/1.fa.fai");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa -r $$opts{tmp}/1.reg -o $$opts{tmp}/out.fa.gz"); #create output for comparison
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/out.fa.gz");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa --write-index -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fa.gz"); #write and index 1.fa.gz.fai and 1.fa.gz.gzi
cmd("$$opts{diff} $$opts{tmp}/out.fa.gz.fai $$opts{tmp}/1.fa.gz.fai");
cmd("$$opts{diff} $$opts{tmp}/out.fa.gz.gzi $$opts{tmp}/1.fa.gz.gzi");
cmd("$$opts{bin}/samtools faidx --fai-idx $$opts{tmp}/fa_test.fai --gzi-idx $$opts{tmp}/fa_test.gzi $$opts{tmp}/faidx.fa.gz -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fa"); #write and index 1.fa.fai
cmd("$$opts{diff} $$opts{tmp}/faidx.fa.fai $$opts{tmp}/1.fa.fai");
cmd("$$opts{bin}/samtools faidx --fai-idx $$opts{tmp}/fa_test.fai --gzi-idx $$opts{tmp}/fa_test.gzi $$opts{tmp}/faidx.fa.gz -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fa.gz"); #write and index 1.fa.gz.fai and 1.fa.gz.gzi
cmd("$$opts{diff} $$opts{tmp}/out.fa.gz.fai $$opts{tmp}/1.fa.gz.fai");
cmd("$$opts{diff} $$opts{tmp}/out.fa.gz.gzi $$opts{tmp}/1.fa.gz.gzi");
# test continuing after an error
cmd("$$opts{bin}/samtools faidx --output $$opts{tmp}/output_faidx.fa --continue $$opts{tmp}/faidx.fa 100 EEE FFF");
# test for reporting retrieval errors, Zero results and truncated
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa 1:10000000-10000005 > $$opts{tmp}/output_faidx.fa 2>&1 && grep Zero $$opts{tmp}/output_faidx.fa");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa 1:99998-100099 > $$opts{tmp}/output_faidx.fa 2>&1 && grep Truncated $$opts{tmp}/output_faidx.fa");
# Get regions from a file
open($fh, ">$$opts{tmp}/region.txt") or error("$$opts{tmp}/region.txt: $!");
print $fh "1\n2:5-10\n3:20-30\n";
close $fh;
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa 1 2:5-10 3:20-30 > $$opts{tmp}/output_faidx_base.fa");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/faidx.fa -r $$opts{tmp}/region.txt > $$opts{tmp}/output_faidx.fa && $$opts{diff} $$opts{tmp}/output_faidx.fa $$opts{tmp}/output_faidx_base.fa");
# reverse complement test
my $fseq = 'ATGAAATGTAACCCAAGAGATATACTCTTCAAGGTACTGTAAGCTATTTCTGTGGACACC';
my $rseq = $fseq;
$rseq =~ tr/ACGTMRWSYKVHDBN/TGCAKYWSRMBDHVN/;
$rseq = reverse $rseq;
open($fh, ">$$opts{tmp}/forward_test.fa") or error("$$opts{tmp}/forward_test.fa: $!");
print $fh ">rc\n$fseq\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test.fa") or error("$$opts{tmp}/rc_answer_test.fa: $!");
print $fh ">rc/rc\n$rseq\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test2.fa") or error("$$opts{tmp}/rc_answer_test2.fa: $!");
print $fh ">rc(-)\n$rseq\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test3.fa") or error("$$opts{tmp}/rc_answer_test3.fa: $!");
print $fh ">rc\n$rseq\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test4.fa") or error("$$opts{tmp}/rc_answer_test3.fa: $!");
print $fh ">rc reverse\n$rseq\n";
close $fh;
cmd("$$opts{bin}/samtools faidx -i $$opts{tmp}/forward_test.fa rc > $$opts{tmp}/forward_test_out.fa && $$opts{diff} $$opts{tmp}/forward_test_out.fa $$opts{tmp}/rc_answer_test.fa");
cmd("$$opts{bin}/samtools faidx --mark-strand sign -i $$opts{tmp}/forward_test.fa rc > $$opts{tmp}/forward_test_out2.fa && $$opts{diff} $$opts{tmp}/forward_test_out2.fa $$opts{tmp}/rc_answer_test2.fa");
cmd("$$opts{bin}/samtools faidx --mark-strand no -i $$opts{tmp}/forward_test.fa rc > $$opts{tmp}/forward_test_out3.fa && $$opts{diff} $$opts{tmp}/forward_test_out3.fa $$opts{tmp}/rc_answer_test3.fa");
cmd("$$opts{bin}/samtools faidx --mark-strand 'custom, forward, reverse' -i $$opts{tmp}/forward_test.fa rc > $$opts{tmp}/forward_test_out4.fa && $$opts{diff} $$opts{tmp}/forward_test_out4.fa $$opts{tmp}/rc_answer_test4.fa");
for my $reg ('3:11-13','2:998-1003','1:100-104','1:99998-100007')
{
for my $file ("$$opts{tmp}/faidx.fa","$$opts{tmp}/faidx.fa.gz")
{
my $test = "$$opts{bin}/samtools faidx $file $reg | grep -v '^>'";
print "$test\n";
my $seq = cmd($test);
chomp($seq);
$seq =~ s/N.*$//;
my $num = faidx_seq_to_num($seq);
my $xreg = $reg;
$xreg =~ s/^[^:]*://;
$xreg =~ s/-.*$//;
if ( $num ne $xreg ) { failed($opts,msg=>$test,reason=>"Expected \"". faidx_num_to_seq($xreg) ."\" got \"$seq\"\n"); }
else { passed($opts,msg=>$test); }
}
}
}
sub fqidx_num_to_qual
{
my ($dec) = @_;
my $out = '';
my @base = qw(I J K L);
while ( $dec>=0 )
{
my $r = $dec % 4;
$out = $base[$r] . $out;
$dec = int( ($dec - $r) / 4 );
if ( !$dec ) { last; }
}
return $out;
}
sub fqidx_qual_to_num
{
my ($seq) = @_;
my $out = 0;
my $len = length($seq);
my %base = ( I=>0, J=>1, K=>2, L=>3 );
for (my $i=0; $i<$len; $i++)
{
my $b = substr($seq,$i,1);
$out += $base{$b} * 4**($len-$i-1);
}
return $out;
}
sub test_fqidx
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
# Create test data: The fake sequence consists of sequence offsets coded
# into A,C,G,T and separated with Ns. The offsets are 1-based.
#
open(my $fh,'>',"$$opts{tmp}/fqidx.fq") or error("$$opts{tmp}/fqidx.fq: $!");
my $ntot = 100_000;
my @dat = qw(A C G T);
for (my $seq=1; $seq<=3; $seq++)
{
my $nwr = 1;
my $out = '';
while ($nwr < $ntot)
{
my $tmp = faidx_num_to_seq($nwr) . 'N';
$out .= $tmp;
$nwr += length($tmp);
}
print $fh "\@$seq\n";
print $fh faidx_wrap($out);
$nwr = 1;
$out = '';
while ($nwr < $ntot) {
my $tmp = fqidx_num_to_qual($nwr) . '!';
$out .= $tmp;
$nwr += length($tmp);
}
print $fh "+\n";
print $fh faidx_wrap($out);
}
close($fh);
# Run tests: index and retrieval from plain text and compressed files
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq");
cmd("cat $$opts{tmp}/fqidx.fq | $$opts{bgzip} -ci -I $$opts{tmp}/fqidx.fq.gz.gzi > $$opts{tmp}/fqidx.fq.gz");
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq.gz");
# Write to file
cmd("$$opts{bin}/samtools fqidx --length 5 $$opts{tmp}/fqidx.fq 1:1-104 > $$opts{tmp}/output_fqidx_base.fq");
cmd("$$opts{bin}/samtools fqidx --length 5 --output $$opts{tmp}/output_fqidx.fq $$opts{tmp}/fqidx.fq 1:1-104 && $$opts{diff} $$opts{tmp}/output_fqidx.fq $$opts{tmp}/output_fqidx_base.fq");
cmd("$$opts{bin}/samtools fqidx --length 5 $$opts{tmp}/fqidx.fq 1 > $$opts{tmp}/output_fqidx_base.1.fq");
# Write to bgzip file
cmd("$$opts{bin}/samtools fqidx --length 5 --output $$opts{tmp}/output_fqidx.fq.1.gz $$opts{tmp}/fqidx.fq 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_fqidx.fq.1.gz && $$opts{diff} $$opts{tmp}/output_fqidx.fq.1 $$opts{tmp}/output_fqidx_base.fq");
cmd("$$opts{bin}/samtools fqidx --length 5 --output $$opts{tmp}/output_fqidx.fq.2.bgz $$opts{tmp}/fqidx.fq 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_fqidx.fq.2.bgz && $$opts{diff} $$opts{tmp}/output_fqidx.fq.2 $$opts{tmp}/output_fqidx_base.fq");
cmd("$$opts{bin}/samtools fqidx --length 5 --output $$opts{tmp}/output_fqidx.fq.3.bgzf $$opts{tmp}/fqidx.fq 1:1-104 && $$opts{bgzip} -df $$opts{tmp}/output_fqidx.fq.3.bgzf && $$opts{diff} $$opts{tmp}/output_fqidx.fq.3 $$opts{tmp}/output_fqidx_base.fq");
# Write to bgzip file with compression level
cmd("$$opts{bin}/samtools fqidx --length 5 --output $$opts{tmp}/output_fqidx.fq.4.gz $$opts{tmp}/fqidx.fq 1 --output-fmt-opt=\"level=4\" && $$opts{bgzip} -df $$opts{tmp}/output_fqidx.fq.4.gz && $$opts{diff} $$opts{tmp}/output_fqidx.fq.4 $$opts{tmp}/output_fqidx_base.1.fq");
# Write to bgzip file with thread
cmd("$$opts{bin}/samtools fqidx ${threads} --length 5 --output $$opts{tmp}/output_fqidx.fq.5.gz $$opts{tmp}/fqidx.fq 1 && $$opts{bgzip} -df $$opts{tmp}/output_fqidx.fq.5.gz && $$opts{diff} $$opts{tmp}/output_fqidx.fq.5 $$opts{tmp}/output_fqidx_base.1.fq");
# Write indices to a file
cmd("$$opts{bin}/samtools fqidx --fai-idx $$opts{tmp}/fq_test.fai --gzi-idx $$opts{tmp}/fq_test.gzi $$opts{tmp}/fqidx.fq.gz");
cmd("$$opts{diff} $$opts{tmp}/fqidx.fq.fai $$opts{tmp}/fq_test.fai");
cmd("$$opts{diff} $$opts{tmp}/fqidx.fq.gz.gzi $$opts{tmp}/fq_test.gzi");
#with write-index
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq 1 --write-index"); #ignores write-index
cmd("echo \"1\n2\n3\" > $$opts{tmp}/1.reg"); #create region file
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq --write-index -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fq"); #writes and indexes
cmd("$$opts{diff} $$opts{tmp}/fqidx.fq.fai $$opts{tmp}/1.fq.fai");
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq -r $$opts{tmp}/1.reg -o $$opts{tmp}/out.fq.gz"); #create output for comparison
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/out.fq.gz");
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq --write-index -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fq.gz"); #write and index 1.fq.gz.fai and 1.fq.gz.gzi
cmd("$$opts{diff} $$opts{tmp}/out.fq.gz.fai $$opts{tmp}/1.fq.gz.fai");
cmd("$$opts{diff} $$opts{tmp}/out.fq.gz.gzi $$opts{tmp}/1.fq.gz.gzi");
cmd("$$opts{bin}/samtools fqidx --fai-idx $$opts{tmp}/fq_test.fai --gzi-idx $$opts{tmp}/fq_test.gzi $$opts{tmp}/fqidx.fq.gz -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fq"); #write and index 1.fa.fai
cmd("$$opts{diff} $$opts{tmp}/fqidx.fq.fai $$opts{tmp}/1.fq.fai");
cmd("$$opts{bin}/samtools fqidx --fai-idx $$opts{tmp}/fq_test.fai --gzi-idx $$opts{tmp}/fq_test.gzi $$opts{tmp}/fqidx.fq.gz -r $$opts{tmp}/1.reg -o $$opts{tmp}/1.fq.gz"); #write and index 1.fa.gz.fai and 1.fa.gz.gzi
cmd("$$opts{diff} $$opts{tmp}/out.fq.gz.fai $$opts{tmp}/1.fq.gz.fai");
cmd("$$opts{diff} $$opts{tmp}/out.fq.gz.gzi $$opts{tmp}/1.fq.gz.gzi");
# test continuing after an error
cmd("$$opts{bin}/samtools fqidx --output $$opts{tmp}/output_fqidx.fq --continue $$opts{tmp}/fqidx.fq 100 EEE FFF");
# test for reporting retrieval errors, Zero results and truncated
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq 1:10000000-10000005 > $$opts{tmp}/output_fqidx.fq 2>&1 && grep Zero $$opts{tmp}/output_fqidx.fq");
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq 1:99998-100099 > $$opts{tmp}/output_fqidx.fq 2>&1 && grep Truncated $$opts{tmp}/output_fqidx.fq");
# Get regions from a file, also test fqidx as faidx -f
open($fh, ">$$opts{tmp}/region.txt") or error("$$opts{tmp}/region.txt: $!");
print $fh "1\n2:5-10\n3:20-30\n";
close $fh;
cmd("$$opts{bin}/samtools fqidx $$opts{tmp}/fqidx.fq 1 2:5-10 3:20-30 > $$opts{tmp}/output_fqidx_base.fq");
cmd("$$opts{bin}/samtools faidx -f $$opts{tmp}/fqidx.fq -r $$opts{tmp}/region.txt > $$opts{tmp}/output_fqidx.fq && $$opts{diff} $$opts{tmp}/output_fqidx.fq $$opts{tmp}/output_fqidx_base.fq");
# reverse complement test
my $fseq = 'ATGAAATGTAACCCAAGAGATATACTCTTCAAGGTACTGTAAGCTATTTCTGTGGACACC';
my $fqual = '0123456789ABCDEFGHIJKLMNOPQRSTUVWXYZabcdefghijklmnopqrstuvwx';
my $rseq = $fseq;
$rseq =~ tr/ACGTMRWSYKVHDBN/TGCAKYWSRMBDHVN/;
$rseq = reverse $rseq;
my $rqual = reverse $fqual;
open($fh, ">$$opts{tmp}/forward_test.fq") or error("$$opts{tmp}/forward_test.fq: $!");
print $fh "\@rc\n$fseq\n+\n$fqual\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test.fq") or error("$$opts{tmp}/rc_answer_test.fq: $!");
print $fh "\@rc/rc\n$rseq\n+\n$rqual\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test2.fq") or error("$$opts{tmp}/rc_answer_test.fq: $!");
print $fh "\@rc\n$rseq\n+\n$rqual\n";
close $fh;
open ($fh, ">$$opts{tmp}/rc_answer_test3.fq") or error("$$opts{tmp}/rc_answer_test.fq: $!");
print $fh "\@rc reverse\n$rseq\n+\n$rqual\n";
close $fh;
cmd("$$opts{bin}/samtools fqidx -i $$opts{tmp}/forward_test.fq rc > $$opts{tmp}/forward_test_out.fq && $$opts{diff} $$opts{tmp}/forward_test_out.fq $$opts{tmp}/rc_answer_test.fq");
cmd("$$opts{bin}/samtools fqidx --mark-strand no -i $$opts{tmp}/forward_test.fq rc > $$opts{tmp}/forward_test_out2.fq && $$opts{diff} $$opts{tmp}/forward_test_out2.fq $$opts{tmp}/rc_answer_test2.fq");
cmd("$$opts{bin}/samtools fqidx --mark-strand 'custom, forward, reverse' -i $$opts{tmp}/forward_test.fq rc > $$opts{tmp}/forward_test_out3.fq && $$opts{diff} $$opts{tmp}/forward_test_out3.fq $$opts{tmp}/rc_answer_test3.fq");
for my $reg ('3:11-13','2:998-1003','1:100-104','1:99998-100007')
{
for my $file ("$$opts{tmp}/fqidx.fq","$$opts{tmp}/fqidx.fq.gz")
{
my $test = "$$opts{bin}/samtools fqidx $file $reg";
print "$test\n";
my $result = cmd($test);
my @fq = split "\n", $result;
my $seq = $fq[1];
$seq =~ s/N.*$//;
my $num = faidx_seq_to_num($seq);
my $xreg = $reg;
$xreg =~ s/^[^:]*://;
$xreg =~ s/-.*$//;
if ( $num ne $xreg ) { failed($opts,msg=>$test,reason=>"Expected \"". faidx_num_to_seq($xreg) ."\" got \"$seq\"\n"); }
else { passed($opts,msg=>$test); }
my $qual = $fq[3];
$qual =~ s/!.*$//;
$num = fqidx_qual_to_num($qual);
if ( $num ne $xreg ) { failed($opts,msg=>$test,reason=>"Expected \"". fqidx_num_to_qual($xreg) ."\" got \"$qual\"\n"); }
else { passed($opts,msg=>$test); }
}
}
}
sub test_dict
{
my ($opts,%args) = @_;
cmd("cat $$opts{path}/dat/dict.fa | $$opts{bgzip} -c > $$opts{tmp}/dict.fa.gz");
test_cmd($opts,out=>'dat/dict.out',cmd=>"$$opts{bin}/samtools dict -a hf37d5 -s 'Homo floresiensis' -u ftp://example.com/hf37d5.fa.gz $$opts{path}/dat/dict.fa");
test_cmd($opts,out=>'dat/dict.out',cmd=>"$$opts{bin}/samtools dict -a hf37d5 -s 'Homo floresiensis' -u ftp://example.com/hf37d5.fa.gz $$opts{tmp}/dict.fa.gz");
test_cmd($opts,out=>'dat/dict.out',cmd=>"cat $$opts{path}/dat/dict.fa | $$opts{bin}/samtools dict -a hf37d5 -s 'Homo floresiensis' -u ftp://example.com/hf37d5.fa.gz");
test_cmd($opts,out=>'dat/dict.alias.out',cmd=>"$$opts{bin}/samtools dict -AH < $$opts{path}/dat/dict.alias.fa");
test_cmd($opts,out=>'dat/dict.alt.out',cmd=>"$$opts{bin}/samtools dict -H -l $$opts{path}/dat/dict.alt < $$opts{path}/dat/dict.alias.fa");
}
sub test_index
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
cmd("$$opts{bin}/samtools view${threads} -b $$opts{path}/dat/large_chrom.sam > $$opts{tmp}/large_chrom.bam");
# test_cmd($opts,out=>'dat/empty.expected',err=>'dat/large_chrom_bai_index.err',cmd=>"$$opts{bin}/samtools index${threads} $$opts{tmp}/large_chrom.bam",want_fail=>1,expect_fail=>1); # command should fail and give an error message, but isn't at the moment
cmd("$$opts{bin}/samtools index${threads} -c $$opts{tmp}/large_chrom.bam");
test_cmd($opts,out=>'dat/large_chrom.out',cmd=>"$$opts{bin}/samtools view${threads} $$opts{tmp}/large_chrom.bam ref2");
test_cmd($opts,out=>'dat/large_chrom.out',cmd=>"$$opts{bin}/samtools view${threads} $$opts{tmp}/large_chrom.bam ref2:1-541556283");
test_cmd($opts,out=>'dat/test_input_1_a.bam.bai.expected',cmd=>"$$opts{bin}/samtools index${threads} $$opts{path}/dat/test_input_1_a.bam && cat $$opts{path}/dat/test_input_1_a.bam.bai",binary=>1);
cmd("$$opts{bin}/samtools index${threads} $$opts{path}/dat/test_input_1_b.bam $$opts{tmp}/test_input_1_b.bam.bai");
test_cmd($opts,out=>'dat/test_input_1_b.X.expected',cmd=>"$$opts{bin}/samtools view${threads} -X $$opts{path}/dat/test_input_1_b.bam $$opts{tmp}/test_input_1_b.bam.bai ref2");
test_cmd($opts,out=>'dat/test_input_1_b.X2.expected',cmd=>"$$opts{bin}/samtools view${threads} -X $$opts{path}/dat/test_input_1_b.bam $$opts{tmp}/test_input_1_b.bam.bai");
test_cmd($opts,out=>'dat/test_input_1_ab.X.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -O sam - -X -cp -R ref2 $$opts{path}/dat/test_input_1_a.bam $$opts{path}/dat/test_input_1_b.bam $$opts{path}/dat/test_input_1_a.bam.bai $$opts{tmp}/test_input_1_b.bam.bai");
# Check -o option
cmd("$$opts{bin}/samtools index${threads} -o $$opts{tmp}/test_input_1_b_opt.bam.bai $$opts{path}/dat/test_input_1_b.bam");
test_cmd($opts,out=>'dat/test_input_1_b.X.expected',cmd=>"$$opts{bin}/samtools view${threads} -X $$opts{path}/dat/test_input_1_b.bam $$opts{tmp}/test_input_1_b_opt.bam.bai ref2");
# Check indexing multiple alignment files
test_cmd($opts,out=>'dat/empty.expected',cmd=>"$$opts{bin}/samtools index${threads} $$opts{path}/dat/test_input_1_a.bam $$opts{path}/dat/test_input_1_b.bam",want_fail=>1);
test_cmd($opts,out=>'dat/test_input_1_a.bam.bai.expected',cmd=>"$$opts{bin}/samtools index${threads} -M $$opts{path}/dat/test_input_1_a.bam $$opts{path}/dat/test_input_1_b.bam && cat $$opts{path}/dat/test_input_1_a.bam.bai",binary=>1);
test_cmd($opts,out=>'dat/test_input_1_b.X.expected',cmd=>"$$opts{bin}/samtools view${threads} -X $$opts{path}/dat/test_input_1_b.bam $$opts{path}/dat/test_input_1_b.bam.bai ref2");
# Check auto-indexing
cmd("$$opts{bin}/samtools view${threads} --write-index -o $$opts{path}/dat/auto_indexed.tmp.bam $$opts{path}/dat/mpileup.1.sam");
test_cmd($opts,out=>"dat/auto_indexed.tmp.bam.csi", cmd=>"$$opts{bin}/samtools index${threads} -c $$opts{path}/dat/auto_indexed.tmp.bam $$opts{tmp}/auto_indexed.csi && cat $$opts{tmp}/auto_indexed.csi", binary=>1);
cmd("$$opts{bin}/samtools view${threads} -T $$opts{path}/dat/mpileup.ref.fa --write-index -o $$opts{path}/dat/auto_indexed.tmp.cram $$opts{path}/dat/mpileup.1.sam");
test_cmd($opts,out=>"dat/auto_indexed.tmp.cram.crai", cmd=>"$$opts{bin}/samtools index${threads} $$opts{path}/dat/auto_indexed.tmp.cram $$opts{tmp}/auto_indexed.crai && cat $$opts{tmp}/auto_indexed.crai", binary=>1);
cmd("$$opts{bin}/samtools view${threads} -h --write-index -O sam,level=5 -o $$opts{path}/dat/auto_indexed.tmp.sam $$opts{path}/dat/mpileup.1.sam");
test_cmd($opts,out=>"dat/auto_indexed.tmp.sam.csi", cmd=>"$$opts{bin}/samtools index${threads} -c $$opts{path}/dat/auto_indexed.tmp.sam $$opts{tmp}/auto_indexed.csi && cat $$opts{tmp}/auto_indexed.csi", binary=>1);
}
sub test_mpileup
{
my ($opts,%args) = @_;
my @files = ('mpileup.1','mpileup.2','mpileup.3');
my $ref = 'mpileup.ref.fa';
# make a local copy, create bams, index the bams and the reference
open(my $fh1,'>',"$$opts{tmp}/mpileup.bam.list") or error("$$opts{tmp}/mpileup.bam.list: $!");
open(my $fh2,'>',"$$opts{tmp}/mpileup.cram.list") or error("$$opts{tmp}/mpileup.cram.list: $!");
open(my $fh3,'>',"$$opts{tmp}/mpileup.bam.urllist") or error("$$opts{tmp}/mpileup.bam.urllist: $!");
open(my $fh4,'>',"$$opts{tmp}/mpileup.cram.urllist") or error("$$opts{tmp}/mpileup.cram.urllist: $!");
for my $file (@files)
{
cmd("$$opts{bin}/samtools view -b $$opts{path}/dat/$file.sam > $$opts{tmp}/$file.bam");
cmd("$$opts{bin}/samtools view -C -T $$opts{path}/dat/mpileup.ref.fa $$opts{path}/dat/$file.sam > $$opts{tmp}/$file.cram");
cmd("$$opts{bin}/samtools index $$opts{tmp}/$file.bam");
cmd("$$opts{bin}/samtools index $$opts{tmp}/$file.cram");
print $fh1 "$$opts{tmp}/$file.bam\n";
print $fh2 "$$opts{tmp}/$file.cram\n";
my $atmp = $^O =~ /^(cygwin|msys)/ ? cygpath($$opts{tmp}) : abs_path($$opts{tmp});
unless ($atmp =~ /^\//) { $atmp = "/$atmp"; }
print $fh3 "file://$atmp/$file.bam\n";
print $fh4 "file://$atmp/$file.cram\n";
}
close($fh1);
close($fh2);
close($fh3);
close($fh4);
cmd("cp $$opts{path}/dat/$ref $$opts{tmp}/$ref");
cmd("$$opts{bgzip} -fi $$opts{tmp}/$ref");
cmd("$$opts{bin}/samtools faidx $$opts{tmp}/$ref.gz");
# print "$$opts{bin}samtools mpileup -gb $$opts{tmp}/mpileup.list -f $$opts{tmp}/$args{ref}.gz > $$opts{tmp}/mpileup.bcf\n";
test_cmd($opts,out=>'dat/mpileup.out.1',err=>'dat/mpileup.err.1',cmd=>"$$opts{bin}/samtools mpileup -b $$opts{tmp}/mpileup.bam.list -f $$opts{tmp}/mpileup.ref.fa.gz -r17:100-150");
test_cmd($opts,out=>'dat/mpileup.out.1',err=>'dat/mpileup.err.1',cmd=>"$$opts{bin}/samtools mpileup -b $$opts{tmp}/mpileup.cram.list -f $$opts{tmp}/mpileup.ref.fa.gz -r17:100-150");
test_cmd($opts,out=>'dat/mpileup.out.1',err=>'dat/mpileup.err.1',cmd=>"$$opts{bin}/samtools mpileup -b $$opts{tmp}/mpileup.bam.urllist -f $$opts{tmp}/mpileup.ref.fa.gz -r17:100-150");
test_cmd($opts,out=>'dat/mpileup.out.1',err=>'dat/mpileup.err.1',cmd=>"$$opts{bin}/samtools mpileup -b $$opts{tmp}/mpileup.cram.urllist -f $$opts{tmp}/mpileup.ref.fa.gz -r17:100-150");
# test that filter mask replaces (not just adds to) default mask
test_cmd($opts,out=>'dat/mpileup.out.3',cmd=>"$$opts{bin}/samtools mpileup -B --ff 0x14 -f $$opts{tmp}/mpileup.ref.fa.gz -r17:1050-1060 $$opts{tmp}/mpileup.1.bam | grep -v mpileup");
test_cmd($opts,out=>'dat/mpileup.out.3',cmd=>"$$opts{bin}/samtools mpileup -B --ff 0x14 -f $$opts{tmp}/mpileup.ref.fa.gz -r17:1050-1060 $$opts{tmp}/mpileup.1.cram | grep -v mpileup");
test_cmd($opts,out=>'dat/mpileup.out.5',cmd=>"$$opts{bin}/samtools mpileup $$opts{path}/mpileup/overlap.bam | grep 128814202");
}
sub test_usage
{
my ($opts,%args) = @_;
my $test = "test_usage";
print "$test:\n";
print "\t$args{cmd}\n";
my $tty_input;
if (-t) {
$args{redirection} = ""; # no redirection necessary
}
elsif (eval { require IO::Pty; $tty_input = new IO::Pty; }) {
# ensure stdin is a terminal, so that subcommands display their usage
$args{redirection} = "<'" . $tty_input->ttyname . "'";
}
else {
warn "$0: no IO::Pty module or can't open pty; skipping usage tests\n";
return;
}
my $command = $args{cmd};
my $commandpath = $$opts{bin}."/".$command;
my ($ret,$out,$err) = _cmd("$commandpath $args{redirection}");
if ( $err =~ m/\/bin\/bash.*no.*such/i ) { failed($opts,msg=>$test,reason=>"could not run $commandpath: $out"); return; }
my @sections = ($err =~ m/(^[A-Za-z]+.*?)(?:(?=^[A-Za-z]+:)|\z)/msg);
my $have_usage = 0;
my $have_version = 0;
my $have_subcommands = 0;
my $usage = "";
my @subcommands = ();
foreach my $section (@sections) {
if ( $section =~ m/^usage/i ) {
$have_usage = 1;
$section =~ s/^[[:word:]]+[[:punct:][:space:]]*//;
$usage = $section;
} elsif ( $section =~ m/^version/i ) {
$have_version = 1;
} elsif ( $section =~ m/^command/i ) {
$have_subcommands = 1;
$section =~ s/^[[:word:]]+[[:punct:]]?[[:space:]]*//;
$section =~ s/^[[:space:]]+//mg;
$section =~ s/^[[:punct:]]+.*?\n//msg;
@subcommands = ($section =~ m/^([[:word:]]+)[[:space:]].*/mg);
}
}
if ( !$have_usage ) { failed($opts,msg=>$test,reason=>"did not have Usage:"); return; }
if ( !$have_version ) { failed($opts,msg=>$test,reason=>"did not have Version:"); return; }
if ( !$have_subcommands ) { failed($opts,msg=>$test,reason=>"did not have Commands:"); return; }
if ( !($usage =~ m/$command/) ) { failed($opts,msg=>$test,reason=>"usage did not mention $command"); return; }
if ( scalar(@subcommands) < 1 ) { failed($opts,msg=>$test,reason=>"could not parse subcommands"); return; }
print "\t$command has subcommands: ".join(", ", @subcommands)."\n";
passed($opts,msg=>$test);
# now test subcommand usage as well
foreach my $subcommand (@subcommands) {
next if ($subcommand =~ /^(help|version)$/);
# Under msys the isatty function fails to recognise the terminal.
# Skip these tests for now.
next if ($^O =~ /^(cygwin|msys)/ && $subcommand =~ /^(dict|sort|stats|view|fasta|fastq|samples|reference|head)$/);
test_usage_subcommand($opts,%args,subcmd=>$subcommand);
}
}
sub test_usage_subcommand
{
my ($opts,%args) = @_;
my $test = "test_usage_subcommand";
print "$test:\n";
print "\t$args{cmd} $args{subcmd}\n";
my $command = $args{cmd};
my $subcommand = $args{subcmd};
my $commandpath = $$opts{bin}."/".$command;
my ($ret,$out,$err) = _cmd("$commandpath $subcommand $args{redirection}");
if ( $err =~ m/\/bin\/bash.*no.*such/i ) { failed($opts,msg=>$test,reason=>"could not run $commandpath $subcommand: $out"); return; }
if ( $err =~ m/not.*implemented/is ) { failed($opts,msg=>$test,reason=>"subcommand indicates it is not implemented",expect_fail=>1); return; }
my $have_usage = 0;
my $usage = "";
my @sections = ($out =~ m/(^[A-Za-z]+.*?)(?:(?=^[A-Za-z]+:)|\z)/msg);
foreach my $section (@sections) {
if ( $section =~ m/^usage/i ) {
$have_usage = 1;
$section =~ s/^[[:word:]]+[[:punct:]]?[[:space:]]*//;
$usage = $section;
}
}
@sections = ($err =~ m/(^[A-Za-z]+.*?)(?:(?=^[A-Za-z]+:)|\z)/msg);
foreach my $section (@sections) {
if ( $section =~ m/^usage/i ) {
$have_usage = 2;
$section =~ s/^[[:word:]]+[[:punct:]]?[[:space:]]*//;
$usage = $section;
}
}
my $fail = 0;
if ( !$have_usage ) { failed($opts,msg=>$test,reason=>"did not have Usage:"); $fail = 1; }
elsif ( $have_usage == 2 ) { failed($opts,msg=>$test,reason=>"Usage on stderr rather than stdout",expect_fail=>1); $fail = 1; }
if ( !($usage =~ m/$command[[:space:]]+$subcommand/) ) { failed($opts,msg=>$test,reason=>"usage did not mention $command $subcommand"); $fail = 1; }
passed($opts,msg=>$test) if $fail == 0;
}
# Add UR tags to @SQ lines in a SAM file.
# UR points to a local file. As this could be anywhere, it is not possible
# to put UR tags into the test SAM files that are checked in to version
# control. Instead this is used to add them when running the tests.
#
# $in is the SAM file to read.
# $out is the SAM file to write.
# $fasta_location is the filename to put in the UR tag.
sub add_ur_tags
{
my ($in, $out, $fasta_location) = @_;
# Add suitable UR: tag to the source SAM file @SQ lines (for CRAM to use)
open(my $sam_in, '<', $in) || die "Couldn't open $in : $!\n";
open(my $sam_out, '>', $out) || die "Couldn't open $out for writing : $!\n";
while (<$sam_in>) {
if (/^\@SQ/) {
chomp;
print $sam_out "$_\tUR:$fasta_location\n"
|| die "Error writing to $out : $!\n";
} else {
print $sam_out $_ || die "Error writing to $out : $!\n";
}
}
close($sam_in) || die "Error reading $in : $!\n";
close($sam_out) || die "Error writing to $out : $!\n";
}
# Calculates the number of reference bases consumed by a CIGAR string.
# $cigar is the CIGAR string.
# returns the number of reference bases.
sub reflen
{
my ($cigar) = @_;
my $len = 0;
my %m = ( M => 1, D => 1, N => 1, '=' => 1, X => 1 );
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) {
if (exists($m{$2})) { $len += $1; }
}
return $len;
}
# Calculates the number of query bases consumed by a CIGAR string.
# $cigar is the CIGAR string.
# returns the number of query bases.
sub querylen
{
my ($cigar) = @_;
my $len = 0;
my %m = ( M => 1, I => 1, S => 1, '=' => 1, X => 1 );
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) {
if (exists($m{$2})) { $len += $1; }
}
return $len;
}
# Filter SAM files.
# This is used to convert a test SAM file into one that has been modified
# in a number of ways. This is then used to compare with the output of
# samtools view, to see if they are the same.
#
# $in is the SAM file to read.
# $out is the SAM file to write.
# $args is a hash ref indicating which filters to apply:
# $args->{no_body} outputs only the header (-H option)
# $args->{no_header} outputs only the alignments (no -h)
# $args->{no_m5} removes M5: tags from header @SQ lines
# $args->{no_sq} removes all @SQ lines from the header
# $args->{min_map_qual} minimum mapping quality it output (-q option)
# $args->{flags_required} bits which must be set in flags (-f option)
# $args->{flags_rejected} bits which must not be set in flags (-F option)
# $args->{read_groups} hash of read groups to output (-r or -R)
# $args->{read_names} names of reads to output (-N)
# $args->{tag} tag used for checking if reads match tag_values (-d or -D)
# $args->{tag_values} hash of values assocated with tag to output (-d or -D)
# $args->{libraries} hash of libraries to output (-l)
# $args->{region} region list to output (-L and region list)
# $args->{strip_tags} hash of tags to strip from alignments (-x)
# $args->{min_qlen} minimum query length to output (-m)
#
# Returns record counts before and after filtering.
#
# The region list is a reference to an array of region definitions. Each
# region definition is itself a reference to an array with between one and
# three elements - reference, start position and end position. The positions
# are inclusive and count from base 1.
#
# * If one element is present, then all the reads aligned to the given
# reference are output.
# * If two elements are present, then all the reads on the given reference
# that end on or after the start position are output.
# * If three elements are present, then all the reads on the given reference
# that end on or after the start position and start on or before the
# end position are output.
#
# N.B. The region list is just a simple filter, so does not work in
# exactly the same way as samtools view which treats the regions as index
# queries. This means samtools view will output the same alignment more than
# once if given overlapping regions, while filter_sam will not. It will
# work in a similar manner to samtools for BED file input though as samtools
# implements BED file regions as a filter rather than a range request.
#
# The BED file support in samtools view is a bit complicated as it actually
# allows two different formats, which can both be mixed in the same file.
#
# The standard BED format has three or more whitespace-separated elements
# on each line, the first three corresponding to reference name, start
# and end positions. Unlike samtools, BED counts bases from zero, and the
# end position is exclusive, i.e. the base after the desired range.
# To convert BED format to a range definition:
# ref S E becomes: ['ref', S + 1, E]
#
# The other format has only two elements (not legal in a normal BED file).
# They define a reference and a single base position. Unfortunately this
# format counts from base one. To convert this to a filter_sam range:
# ref P becomes: ['ref', P, P]
sub filter_sam
{
my ($in, $out, $args) = @_;
my $no_body = exists($args->{no_body});
my $no_header = exists($args->{no_header});
my $no_m5 = exists($args->{no_m5});
my $no_sq = exists($args->{no_sq});
my $min_map_qual = $args->{min_map_qual} || 0;
my $flags_required = $args->{flags_required} || 0;
my $flags_rejected = $args->{flags_rejected} || 0;
my $read_groups = $args->{read_groups};
my $read_names = $args->{read_names};
my $tag = $args->{tag};
my $tag_values = $args->{tag_values};
my $libraries = $args->{libraries};
my $region = $args->{region};
my $strip_tags = $args->{strip_tags};
my $min_qlen = $args->{min_qlen} || 0;
my $body_filter = ($flags_required || $flags_rejected
|| $read_groups || $read_names || $tag_values
|| $min_map_qual || $libraries || $region
|| $strip_tags || $min_qlen);
my $lib_read_groups = $libraries ? {} : undef;
open(my $sam_in, '<', $in) || die "Couldn't open $in : $!\n";
open(my $sam_out, '>', $out) || die "Couldn't open $out for writing : $!\n";
my $total_records = 0;
my $output_records = 0;
while (<$sam_in>) {
chomp;
if (/^@/) {
next if ($no_header);
if ($libraries && /^\@RG/) {
my ($id) = /\tID:([^\t]+)/;
my ($lib) = /\tLB:([^\t]+)/;
if (exists($libraries->{$lib||""})) {
$lib_read_groups->{$id} = 1;
}
}
if ($read_groups && /^\@RG/) {
my ($id) = /\tID:([^\t]+)/;
next if (!exists($read_groups->{$id||""}));
}
next if ($no_sq && /^\@SQ/);
if ($no_m5 && /^\@SQ/) {
s/\tM5:[^\t\n]+//;
}
print $sam_out "$_\n" || die "Error writing to $out : $!\n";
} else {
next if ($no_body);
$total_records++;
if ($body_filter) {
my @sam = split(/\t/, $_);
next if ($flags_required
&& ($sam[1] & $flags_required) != $flags_required);
next if ($flags_rejected && ($sam[1] & $flags_rejected) != 0);
next if ($min_map_qual && $sam[4] < $min_map_qual);
if ($read_groups || $lib_read_groups) {
my $group = '';
for my $i (11 .. $#sam) {
last if (($group) = $sam[$i] =~ /^RG:Z:(.*)/);
}
next if ($read_groups && !exists($read_groups->{$group||""}));
next if ($lib_read_groups
&& !exists($lib_read_groups->{$group||""}));
}
if ($tag_values) {
my $tag_value = '';
for my $i (11 .. $#sam) {
last if (($tag_value) = $sam[$i] =~ /^${tag}:[ZiIsScCA]:(.*)/);
}
next if (!exists($tag_values->{$tag_value||""}));
}
if ($read_names) {
next if (!exists($read_names->{$sam[0]}));
}
if ($region) {
my $in_range = 0;
foreach my $r (@$region) {
next if ($r->[0] ne $sam[2]);
next if (@$r > 1
&& $sam[3] + reflen($sam[5]) - 1 < $r->[1]);
next if (@$r > 2 && $sam[3] > $r->[2]);
$in_range = 1;
last;
}
next if (!$in_range);
}
if ($min_qlen > 0) {
next if (querylen($sam[5]) < $min_qlen);
}
if ($strip_tags) {
my $stripped = 0;
for (my $i = $#sam; $i >= 11; --$i) {
if (exists($strip_tags->{substr($sam[$i], 0, 2)})) {
$stripped = 1;
splice(@sam, $i, 1);
}
}
if ($stripped) { $_ = join("\t", @sam); }
}
}
$output_records++;
print $sam_out "$_\n" || die "Error writing to $out : $!\n";
}
}
close($sam_in) || die "Error reading $in : $!\n";
close($sam_out) || die "Error writing to $out : $!\n";
return ($total_records, $output_records);
}
# Run a samtools subcommand test. As well as running samtools, various
# options are available to compare the output with a known version. The
# default subcommand run is 'view', unless overridden by the 'cmd' argument.
# This will call passed() or failed() depending on the outcome of the test.
#
# $opts is the global settings hash
# %args is a list of key => value pairs giving options for run_view_test.
# The options for running samtools view are:
# msg => Message describing the test.
# cmd => samtools subcommand to run (default is view)
# args => array ref of arguments to add to the samtools view command
# Does not include the output file, which is in $args{out}
# stdin => Redirect STDIN to a pipe running "cat $args{stdin}"
# Otherwise the input file is included in $args{args}
# out => Name of the output file to make.
# redirect => If set, redirect STDOUT, otherwise use "-o $args{out}"
# ref_path => Setting for the REF_PATH environment variable
# expect_fail => Expected failure, convert failed to xfail and passed to xpass
#
# One of the compare* options can be used to compare to an existing file:
# compare => Compare $args{out} with $args{compare} using sam_compare()
# compare_sam => As compare, but run "samtools view -h $args{out}" first to
# convert an alternate format to SAM. The resulting SAM file
# is saved to "$args{out}.sam" unless $args{pipe} is set.
# pipe => When used with compare_sam, the "samtools view -h" is read
# from a pipe instead of being stored in a file. This can
# be used to avoid making very big SAM files for some tests.
# compare_bam => Compare $args{out} with $args{compare_bam} using bam_compare()
# This is a binary comparison of the gunzipped BAM files.
# compare_count => Compare count stored in $args{out} and $args{count_compare}
# The latter can either be a number, or a file containing a
# number.
# compare_text => Compare $args{out} with $args{compare_text} using
# text_compare(). This is an exact line-by-line comparison.
sub run_view_test
{
my ($opts, %args) = @_;
printf "\t%-60s", $args{msg};
local $ENV{REF_PATH} = $args{ref_path} if ($args{ref_path});
my @cmd = ("$$opts{bin}/samtools", $args{cmd} ? $args{cmd} : 'view');
if ($args{out} && !$args{redirect}) { push(@cmd, '-o', $args{out}); }
if ($args{args}) { push(@cmd, @{$args{args}}); }
my $pid = fork();
unless (defined($pid)) { die "Couldn't fork : $!\n"; }
my $save_stdout;
if ($^O eq 'MSWin32') {
# Ensure we can restore STDOUT properly on Windows. Not doing this
# causes output to be lost if STDOUT is redirected below.
open($save_stdout, '>&STDOUT') || die "Couldn't dup STDOUT: $!\n";
}
unless ($pid) {
if ($args{stdin}) {
open(STDIN, '-|', 'cat', $args{stdin})
|| die "Couldn't pipe cat $args{stdin} to STDIN : $!\n";
}
if ($args{redirect} && $args{out}) {
open(STDOUT, '>', $args{out})
|| die "Couldn't redirect STDOUT to $args{out} : $!\n";
}
exec(@cmd) || die "Couldn't exec @cmd : $!\n";
}
my $reaped = waitpid($pid, 0);
my $res = $reaped == $pid && $? == 0 ? 0 : 1;
if ($^O eq 'MSWin32') {
open(STDOUT, '>&', $save_stdout)
|| die "Couldn't restore STDOUT : $!\n";
}
if (!$res && $args{compare_sam} && $args{out}) {
# Convert output back to sam and compare
my $sam_name = "$args{out}.sam";
my @cmd2 = ("$$opts{bin}/samtools", 'view', '-h', '--no-PG');
if (!$args{pipe}) {
push(@cmd2, '-o', $sam_name);
}
push(@cmd2, $args{out});
push(@cmd, '&&', @cmd2); # For the 'Failed command' message below
my $sam_out;
if (!$args{pipe}) {
$sam_out = $sam_name;
$res = system(@cmd2) == 0 ? 0 : 1;
} else {
open($sam_out, '-|', @cmd2)
|| die "Couldn't open pipe from @cmd2: $!\n";
binmode($sam_out);
}
# Hack $args so the comparison gets done
$args{compare} = $args{compare_sam};
$args{out} = $sam_out;
}
if (!$res) {
if ($args{compare} && $args{out}) {
$res = sam_compare($opts, $args{out}, $args{compare});
} elsif ($args{compare_bam} && $args{out}) {
$res = bam_compare($opts, $args{out}, $args{compare_bam});
} elsif ($args{compare_count}) {
$res = count_compare($args{out}, $args{compare_count});
} elsif ($args{compare_text}) {
$res = text_compare($args{out}, $args{compare_text});
}
}
if (!$res && $args{check_save_counts}) {
$res = check_saved_counts(@{$args{check_save_counts}});
}
if ($res) {
print "\tFailed command:\n\t@cmd\n\n";
failed($opts, %args);
} else {
passed($opts, %args);
}
}
# Check the contents of a file made by the view --save-counts option
sub check_saved_counts {
my ($counts_file, $total, $accepted) = @_;
local $/ = undef;
open(my $in, '<', $counts_file) || die "Couldn't open $counts_file $!\n";
my $content = <$in>;
close($in) || die "Error reading $counts_file $!\n";
$content =~ s/\r\n/\n/g;
my ($file_total, $file_accepted, $file_rejected, $trailing_content) =
$content =~ m/\{\n
\s+"records_processed"\ :\ (\d+),\n
\s+"records_filter_accepted"\ :\ (\d+),\n
\s+"records_filter_rejected"\ :\ (\d+)\n
\}\n(.*)/xs;
my $ok = (defined($file_total) && $file_total == $total
&& defined($file_accepted) && $file_accepted == $accepted
&& defined($file_rejected) && $file_rejected == $total - $accepted
&& !$trailing_content);
if (!$ok) {
printf(STDERR "\nSaved counts file content differs. Got:\n%s\n"
. "Expected:\n"
. "{\n"
. " \"records_processed\" : %d,\n"
. " \"records_filter_accepted\" : %d,\n"
. " \"records_filter_rejected\" : %d\n"
. "}\n", $content, $total, $accepted, $total - $accepted);
return 1;
}
return 0;
}
# Runs a test of the samtools view -s subsampling option.
# The subsampling is pseudo-random, so multiple tests are run with different
# seeds, and the resulting read counts are tested against an acceptable
# range. As the input file is paired, this also checks that both reads from
# each pair get returned.
#
# $opts is the global settings hash
# %args is a list of key => value pairs giving options
# The options are:
# msg => Message describing the test.
# trials => The number of trials to run. The trial number is used as the seed.
# input => The input file to read.
# frac => The fraction of reads to return (must be < 1).
# min => The minimum acceptable read count
# max => The maximum acceptable read count
sub run_view_subsample_test
{
my ($opts, %args) = @_;
printf "\t%s ", $args{msg};
my @counts;
my $res = 0;
for (my $try = 0; $res == 0 && $try < $args{trials}; $try++) {
my %reads;
my @cmd = ("$$opts{bin}/samtools", 'view',
'-s', $try + $args{frac}, $args{input});
open(my $samp, '-|', @cmd) || die "Couldn't open pipe from @cmd: $!\n";
while (<$samp>) {
my ($name) = split(/\t/);
$reads{$name}++;
}
close($samp) || die "Error running @cmd\n";
my $count = 0;
while (my ($name, $num) = each %reads) {
if ($num != 2) {
print "\n\tGot one of read $name, expected two.\n";
$res = 1;
last;
}
$count += $num;
}
print ".";
push(@counts, $count);
}
if (0 == @counts) {
print "samtools view -s returned no counts\n";
$res = 1;
}
@counts = sort { $a <=> $b } @counts;
if ($counts[0] < $args{min} || $counts[-1] > $args{max}) {
printf("\n\tOutput out of range: target (%d..%d) got (%d..%d)\n",
$args{min}, $args{max}, $counts[0], $counts[-1]);
$res = 1;
}
if (!$res) {
passed($opts, %args);
return;
}
failed($opts, %args);
}
# Open a pipe from bgzip to decompress a gzipped file. bgzip is annoying
# as it checks the suffix of the file it is decompressing even when
# writing to STDOUT. Hence we get it to read from a redirected STDIN
# instead so it doesn't care about the filename any more.
#
# $opts is the global settings hash
# $in is the compressed file to read
#
# Returns a file handle reference to the pipe from bgzip.
sub open_bgunzip
{
my ($opts, $in) = @_;
my $bgzip;
open($bgzip, "$$opts{bgzip} -c -d < $in |")
|| die "Couldn't open pipe to bgzip -c -d $!\n";
binmode($bgzip);
return $bgzip;
}
# Compare two SAM files. Headers are collated by type to allow for reordering
# although all headers of a particular type should be in the same order.
# Optional tags on alignment lines can also be reordered.
#
# $opts is the global settings hash
# $sam1 is the first SAM file
# $sam2 is the second SAM file.
#
# Returns 0 if the files were the same, 1 if different.
#
# If $sam1 is a reference, it is assumed to be a pipe from which the data
# should be read. The pipe will be closed after being read. Otherwise it
# is treated as a filename.
#
# If $sam2 ends in '.gz' it is uncompressed with bgzip.
sub sam_compare
{
my ($opts, $sam1, $sam2) = @_;
unless (-e $sam2) {
print "\n\tMissing SAM file $sam2.\n";
return 1;
}
unless ((ref($sam1) || -e $sam1)) {
print "\n\tMissing SAM file $sam1.\n";
return 1;
}
my %hdr1;
my %hdr2;
my ($lno1, $lno2) = (0, 0);
my ($l1, $l2, $ht1, $ht2);
my $f1;
if (ref($sam1)) {
$f1 = $sam1;
} else {
open($f1, '<', $sam1) || die "Couldn't open $sam1: $!\n";
}
while ($l1 = <$f1>) {
$lno1++;
if (($ht1) = $l1 =~ /^(@\S+)/) {
$l1 =~ s/\x0d\x0a$/\x0a/;
push(@{$hdr1{$ht1}}, $l1);
} else {
last;
}
}
my $f2;
if ($sam2 =~ /\.gz$/) {
$f2 = open_bgunzip($opts, $sam2);
} else {
open($f2, '<', $sam2) || die "Couldn't open $sam2: $!\n";
}
while ($l2 = <$f2>) {
$lno2++;
if (($ht2) = $l2 =~ /^(@\S+)/) {
$l2 =~ s/\x0d\x0a$/\x0a/;
push(@{$hdr2{$ht2}}, $l2);
} else {
last;
}
}
while (my ($ht, $h1) = each %hdr1) {
my $h2 = $hdr2{$ht};
my $same = $h2 && @$h1 == @$h2;
if ($same) {
for (my $i = 0; $i < @$h1; $i++) {
next if ($h1->[$i] eq $h2->[$i]);
# Hack to deal with CRAM adding M5 tags
if ($ht eq '@SQ' && $h1->[$i] =~ /\tM5/ && $h2->[$i] !~ /\tM5/){
$h1->[$i] =~ s/\tM5:[0-9a-f]+//;
next if ($h1->[$i] eq $h2->[$i]);
}
# Hack to handle PP, CL and VN tags for @PG lines
if ($ht eq '@PG') {
$h1->[$i] =~ s/\tVN:[^\t\n]+//;
$h2->[$i] =~ s/\tVN:[^\t\n]+//;
$h1->[$i] =~ s/\tCL:[^\t\n]+//;
$h2->[$i] =~ s/\tCL:[^\t\n]+//;
$h1->[$i] =~ s/\tPP:[^\t\n]+//;
$h2->[$i] =~ s/\tPP:[^\t\n]+//;
next if ($h1->[$i] eq $h2->[$i]);
}
$same = 0;
last;
}
}
if (!$same) {
print "\n\tHeader type $ht differs.\n";
print "\t$sam1 has:\n";
foreach my $t (@{$hdr1{$ht}}) {
my $n = length($t);
print "\t|$t| ($n)\n";
}
print "\t$sam2 has:\n";
foreach my $t (@{$hdr2{$ht}}) {
my $n = length($t);
print "\t|$t| ($n)\n";
}
close($f1);
close($f2);
return 1;
}
}
while ($l1 && $l2) {
chomp($l1); $l1 =~ s/\r$//;
chomp($l2); $l2 =~ s/\r$//;
my @sam1 = split(/\t/, $l1);
my @sam2 = split(/\t/, $l2);
my @tags1 = sort(splice(@sam1, 11));
my @tags2 = sort(splice(@sam2, 11));
# Windows uses e.g. 1.9e+009 vs 1.9e+09 on Linux
@tags1 = map {s/(e[-+])0(\d\d)/$1$2/g;$_} @tags1;
@tags2 = map {s/(e[-+])0(\d\d)/$1$2/g;$_} @tags2;
if (join("\t", @sam1, @tags1) ne join("\t", @sam2, @tags2)) {
last;
}
$l1 = <$f1>;
$l2 = <$f2>;
$lno1++;
$lno2++;
}
close($f1) || die "Error reading $sam1: $!\n";
close($f2) || die "Error reading $sam2: $!\n";
if ($l1 || $l2) {
print "\n"; STDOUT->flush();
print STDERR "\tSAM files differ at $sam1 line $lno1 / $sam2 line $lno2\n";
print STDERR "$l1\n$l2\n";
return 1;
}
return 0;
}
# Compare two BAM files. This is done by uncompressing them and performing
# a binary comparison. Used to check that (for example) the uncompressed and
# compressed versions are the same.
#
# $opts is the global settings hash
# $bam1 is the first BAM file
# $bam2 is the second BAM file.
#
# Returns 0 if the files are the same, 1 if they are different.
sub bam_compare
{
my ($opts, $bam1, $bam2) = @_;
my $buffer1;
my $buffer2;
my $bytes1;
my $bytes2;
my $fail = 0;
my $b1 = open_bgunzip($opts, $bam1);
my $b2 = open_bgunzip($opts, $bam2);
do {
$bytes1 = read($b1, $buffer1, 65536);
$bytes2 = read($b2, $buffer2, 65536);
if (!defined($bytes1)) { die "Error reading $bam1 : $!\n"; }
if (!defined($bytes2)) { die "Error reading $bam2 : $!\n"; }
if ($bytes1 != $bytes2 || $buffer1 ne $buffer2) {
$fail = 1;
}
} while ($bytes1 && $bytes2 && !$fail);
close($b1) || die "Error running bgzip -c -d $bam1\n";
close($b2) || die "Error running bgzip -c -d $bam2\n";
if ($fail) {
print "\n\tBAM files $bam1 and $bam2 differ.\n";
return 1;
}
return 0;
}
# Compare two text files.
#
# $txt1 is the first file
# $txt2 is the second file.
#
# Returns 0 if the files are the same, 1 if they are different.
sub text_compare
{
my ($txt1, $txt2) = @_;
open(my $t1, '<', $txt1) || die "Couldn't open $txt1 : $!\n";
open(my $t2, '<', $txt2) || die "Couldn't open $txt2 : $!\n";
my $line = 0;
my $diff = 0;
while (!$diff) {
$line++;
my $l1 = <$t1>;
my $l2 = <$t2>;
last if (!defined($l1) && !defined($l2));
$l1 =~ s/\x0d\x0a$/\x0a/;
$l2 =~ s/\x0d\x0a$/\x0a/;
if (($l1 || '') ne ($l2 || '')) {
$diff = 1;
if (defined($l1)) { chomp($l1); }
if (defined($l2)) { chomp($l2); }
print "\n\tFiles differ at line $line:\n";
print "\t$txt1 ", defined($l1) ? " : $l1\n" : "End of file\n";
print "\t$txt2 ", defined($l2) ? " : $l2\n" : "End of file\n";
}
}
close($t1) || die "Error reading $t1 : $!\n";
close($t2) || die "Error reading $t2 : $!\n";
return $diff;
}
# Compare two counts to see if they are the same. The first is in a file
# created by, for example, 'samtools view -c'. The second can either be
# a number, or the name of a file to read.
#
# $count1 is the first file containing a count
# $count2 is either a number or the second file containing a count
#
# Returns 0 if the counts were the same, 1 if they differ.
sub count_compare
{
my ($count1, $count2) = @_;
open(my $c1, '<', $count1) || die "Couldn't open $count1 : $!\n";
my $number1 = <$c1>;
chomp($number1); $number1 =~ s/\r$//;
close($c1) || die "Error reading $count1 : $!\n";
unless ($number1 =~ /^\d+$/) {
print "\n\tExpected a number in $count1 but got '$number1'\n";
return 1;
}
my $number2 = 0;
if ($count2 =~ /^\d+$/) {
$number2 = $count2;
} else {
open(my $c2, '<', $count2) || die "Couldn't open $count2 : $!\n";
while (<$c2>) {
if (!/^@/) { $number2++; }
}
close($c2) || die "Error reading $count2 : $!\n";
}
if ($number1 != $number2) {
print "\n\tIncorrect count: Got $number1; expected $number2.\n";
return 1;
}
return 0;
}
# Generate a pair of reads, for use in gen_file.
#
# $reads is a hash that the reads are put into.
# $seq is the sequence (passed by reference for speed)
# $qual is the quality (passed by reference for speed)
# $pos1 is the position of the leftmost read.
# $size is the length of $$seq
# $rnum is the read number, used to generate the read name.
sub gen_pair
{
my ($reads, $seq, $qual, $pos1, $size, $rnum) = @_;
my $l1 = int(rand(50) + 75);
my $l2 = int(rand(50) + 75);
my $pos2 = $pos1 + int(rand(50) + 275);
if ($pos2 + $l2 > $size) { return; }
my $orient = int(rand(2));
my $name = "ERR123456.${rnum}";
my $rd1 = sprintf("%s\t%d\tref1\t%d\t40\t%dM\t=\t%d\t%d\t%s\t%s\tNM:i:0\tMD:Z:$l1\tRG:Z:g1",
$name, $orient ? 99 : 163, $pos1 + 1, $l1, $pos2 + 1,
$pos2 + $l2 - $pos1, substr($$seq, $pos1, $l1),
substr($$qual, $pos1, $l1));
my $rd2 = sprintf("%s\t%d\tref1\t%d\t40\t%dM\t=\t%d\t%d\t%s\t%s\tNM:i:0\tMD:Z:$l2\tRG:Z:g1",
$name, $orient ? 147 : 83, $pos2 + 1, $l2, $pos1 + 1,
-($pos2 + $l2 - $pos1), substr($$seq, $pos2, $l2),
substr($$qual, $pos2, $l2));
push(@{$reads->{$pos1}}, $rd1);
push(@{$reads->{$pos2}}, $rd2);
}
# Randomly generate a test SAM file. Used to create the big SAM file for
# testing the -s and -@ options of samtools view. As well as the SAM file
# it writes a fasta file with the reference sequence in it and a .fai file
# which is an index of the fasta file. Both of these are used for CRAM.
# The SAM file is compressed with bgzf to keep the size down.
#
# To generate random sequence aligned against identical referenes,
# pass in the 4th argument (ref_seed) with a constant value. Otherwise a
# random reference is generated too.
#
# $opts is the global setting hash.
# $prefix is the prefix for the generated file names.
# $size is the length of the random reference sequence.
#
# Returns a list containing the name of the SAM file and the read count.
sub gen_file
{
my ($opts, $prefix, $size, $ref_seed) = @_;
local $| = 1;
$ref_seed = rand(1<<31) unless defined($ref_seed);
my $pair_seed = rand(1<<31);
print "\tGenerating test data file ";
my $dot_interval = $size / 10;
my $seq = "!" x $size;
my $qual = $seq;
srand($ref_seed);
my $next_dot = $dot_interval;
for (my $b = 0; $b < $size; $b++) {
if ($b == $next_dot) {
print ".";
$next_dot += $dot_interval;
}
substr($seq, $b, 1) = (qw(A C G T))[int(rand(4))];
substr($qual, $b, 1) = chr(33 + int(rand(40)));
}
my $fasta = "$prefix.fa";
open(my $fa, '>', $fasta) || die "Couldn't open $fasta for writing : $!\n";
binmode($fa);
print $fa ">ref1\n";
for (my $i = 0; $i < $size; $i += 60) {
print $fa substr($seq, $i, 60), "\n";
}
close($fa) || die "Error writing to $fasta : $!\n";
my $fai = "$prefix.fa.fai";
open($fa, '>', $fai) || die "Couldn't open $fai for writing : $!\n";
binmode($fa);
print $fa "ref1\t$size\t6\t60\t61\n";
close($fa) || die "Error writing to $fai : $!\n";
my $sam = "$prefix.sam.gz";
open(my $s, "| $$opts{bgzip} -c > $sam")
|| die "Couldn't open pipe to bgzip $!";
binmode($s);
print $s "\@HD\tVN:1.4\tSO:coordinate\n";
print $s "\@RG\tID:g1\tDS:Group 1\tLB:Lib1\tSM:Sample1\n";
print $s "\@SQ\tSN:ref1\tLN:$size\tUR:$fasta\n";
my %read_store;
my $rnum = 0;
$next_dot = $dot_interval;
srand($pair_seed);
for (my $i = 0; $i < $size; $i++) {
if ($i == $next_dot) {
print ".";
$next_dot += $dot_interval;
}
if ($i % 20 == 0) {
gen_pair(\%read_store, \$seq, \$qual, $i, $size, ++$rnum);
}
if (exists($read_store{$i})) {
foreach my $read (@{$read_store{$i}}) {
print $s $read, "\n";
}
delete($read_store{$i});
}
}
close($s) || die "Error running bgzip -c > $sam : $!\n";
print " done\n";
return ($sam, $rnum * 2);
}
# Run samtools view tests.
sub test_view
{
my ($opts) = @_;
my $test_name = "test_view";
print "$test_name:\n";
my $ref_path = "$$opts{path}/dat/cram_md5/%s";
# Add @SQ UR: tags to the stored test file.
my $sam_no_ur = "$$opts{path}/dat/view.001.sam";
my $sam_with_ur = "$$opts{tmp}/view.001.sam";
add_ur_tags($sam_no_ur, $sam_with_ur, "$$opts{path}/dat/view.001.fa");
# Generate a big SAM file for use later.
my ($big_sam, $big_sam_count)
= gen_file($opts, "$$opts{tmp}/view.big", 1000000);
my $test = 1;
my $out = "$$opts{tmp}/view";
# SAM -> BAM -> SAM
my $bam_with_ur_out = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> BAM -> SAM",
args => ['-b', $sam_with_ur, '--no-PG'],
out => $bam_with_ur_out,
compare_sam => $sam_with_ur);
$test++;
# SAM -> uncompressed BAM -> SAM
my $uncomp_bam = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> uncompressed BAM",
args => ['-u', $sam_with_ur, '--no-PG'],
out => $uncomp_bam,
compare_bam => $bam_with_ur_out);
$test++;
run_view_test($opts,
msg => "$test: uncompressed BAM -> SAM and compare",
args => ['-h', $uncomp_bam, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_with_ur);
$test++;
# SAM -> fast compressed BAM -> SAM
my $fastcomp_bam = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> fast compressed BAM",
args => ['-1', $sam_with_ur, '--no-PG'],
out => $fastcomp_bam,
compare_bam => $bam_with_ur_out);
$test++;
run_view_test($opts,
msg => "$test: fast compressed BAM -> SAM and compare",
args => ['-h', $fastcomp_bam, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_with_ur);
$test++;
# SAM -> CRAM -> SAM with UR tags
my $cram_with_ur_out = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> CRAM -> SAM (UR header tags)",
args => ['-C', $sam_with_ur, '--no-PG'],
out => $cram_with_ur_out,
compare_sam => $sam_with_ur);
$test++;
# SAM -> CRAM -> SAM with M5 tags
run_view_test($opts,
msg => "$test: SAM -> CRAM -> SAM (M5 header tags)",
args => ['-C', $sam_no_ur, '--no-PG'],
out => sprintf("%s.test%03d.cram", $out, $test),
compare_sam => $sam_no_ur,
ref_path => $ref_path);
$test++;
# SAM -> BAM -> CRAM -> SAM with UR tags
my $cram_from_bam = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: BAM -> CRAM with UR -> SAM",
args => ['-C', $bam_with_ur_out, '--no-PG'],
out => $cram_from_bam,
compare_sam => $sam_with_ur);
$test++;
# SAM -> BAM -> CRAM -> SAM with M5 tags
my $bam_no_ur = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> BAM (M5 header tags)",
args => ['-b', $sam_no_ur, '--no-PG'],
out => $bam_no_ur,
compare_sam => $sam_no_ur);
$test++;
my $cram_no_ur = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> BAM -> CRAM -> SAM (M5 header tags)",
args => ['-C', $bam_no_ur, '--no-PG'],
out => $cram_no_ur,
compare_sam => $sam_no_ur,
ref_path => $ref_path);
$test++;
# SAM -> BAM -> CRAM -> BAM -> SAM with UR tags
my $bam_from_cram = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: CRAM -> BAM with UR",
args => ['-b', $cram_from_bam, '--no-PG'],
out => $bam_from_cram,
compare_sam => $sam_with_ur);
$test++;
# SAM -> BAM -> CRAM -> BAM -> SAM with M5 tags
run_view_test($opts,
msg => "$test: CRAM -> BAM with M5",
args => ['-b', $cram_no_ur, '--no-PG'],
out => sprintf("%s.test%03d.bam", $out, $test),
compare_sam => $sam_no_ur,
ref_path => $ref_path);
$test++;
# Write to stdout
run_view_test($opts,
msg => "$test: SAM -> SAM via stdout",
args => ['-h', $sam_with_ur, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
redirect => 1,
compare => $sam_with_ur);
$test++;
run_view_test($opts,
msg => "$test: SAM -> BAM via stdout",
args => ['-b', $sam_with_ur, '--no-PG'],
out => sprintf("%s.test%03d.bam", $out, $test),
redirect => 1,
compare_bam => $bam_with_ur_out);
$test++;
my $cram_via_stdout = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: SAM -> CRAM via stdout",
args => ['-C', $sam_with_ur, '--no-PG'],
out => $cram_via_stdout,
redirect => 1,
compare_sam => $sam_with_ur);
$test++;
# Read from stdin
run_view_test($opts,
msg => "$test: SAM from stdin -> SAM",
args => ['-h', '-', '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
stdin => $sam_with_ur,
compare => $sam_with_ur);
$test++;
run_view_test($opts,
msg => "$test: BAM from stdin -> SAM",
args => ['-h', '-', '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
stdin => $bam_with_ur_out,
compare => $sam_with_ur);
$test++;
run_view_test($opts,
msg => "$test: CRAM from stdin -> SAM",
args => ['-h', '-', '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
stdin => $cram_with_ur_out,
compare => $sam_with_ur);
$test++;
# Header only options
my $sam_header = "$$opts{tmp}/view.001.header.sam";
filter_sam($sam_with_ur, $sam_header, {no_body => 1});
run_view_test($opts,
msg => "$test: samtools view -H (SAM input)",
args => ['-H', $sam_with_ur, '--no-PG'],
out => sprintf("%s.test%03d.header", $out, $test),
compare => $sam_header);
$test++;
run_view_test($opts,
msg => "$test: samtools view -H (BAM input)",
args => ['-H', $bam_with_ur_out, '--no-PG'],
out => sprintf("%s.test%03d.header", $out, $test),
compare => $sam_header);
$test++;
run_view_test($opts,
msg => "$test: samtools view -H (CRAM input)",
args => ['-H', $cram_with_ur_out, '--no-PG'],
out => sprintf("%s.test%03d.header", $out, $test),
compare => $sam_header);
$test++;
# Body only
my $sam_body = "$$opts{tmp}/view.001.body.sam";
filter_sam($sam_with_ur, $sam_body, {no_header => 1});
run_view_test($opts,
msg => "$test: No headers (SAM input)",
args => [$sam_with_ur, '--no-PG'],
out => sprintf("%s.test%03d.body", $out, $test),
compare => $sam_body);
$test++;
run_view_test($opts,
msg => "$test: No headers (BAM input)",
args => [$bam_with_ur_out, '--no-PG'],
out => sprintf("%s.test%03d.body", $out, $test),
compare => $sam_body);
$test++;
run_view_test($opts,
msg => "$test: No headers (CRAM input)",
args => [$cram_with_ur_out, '--no-PG'],
out => sprintf("%s.test%03d.body", $out, $test),
compare => $sam_body);
$test++;
# Filter and counting tests.
# Group names file for -R test
my $fogn = "$$opts{tmp}/view.001.fogn";
open(my $f, '>', $fogn) || die "Couldn't open $fogn : $!\n";
print $f "grp1\ngrp3\n" || die "Error writing to $fogn : $!\n";
close($f) || die "Error writing to $fogn : $!\n";
# Barcodes file for -D test
my $fobc = "$$opts{tmp}/view.001.fobc";
open($f, '>', $fobc) || die "Couldn't open $fobc : $!\n";
print $f "ACGT\nAATTCCGG\n" || die "Error writing to $fobc : $!\n";
close($f) || die "Error writing to $fobc : $!\n";
# Read names file for -N test
my $forn = "$$opts{tmp}/view.001.forn";
open($f, '>', $forn) || die "Couldn't open $forn : $!\n";
print $f "ref1_grp1_p001\nunaligned_grp3_p001\nr008\nr009\n" || die "Error writing to $forn : $!\n";
close($f) || die "Error writing to $forn : $!\n";
my @filter_tests = (
# [test_name, {filter_sam options}, [samtools options], expect_fail]
# Flags
['req128', {flags_required => 128}, ['-f', 128], 0],
['rej128', {flags_rejected => 128}, ['-F', 128], 0],
['rej128req2', { flags_rejected => 128, flags_required => 2 },
['-F', 128, '-f', 2], 0],
# Read groups
['rg_grp2', { read_groups => { grp2 => 1 }}, ['-r', 'grp2'], 0],
['rg_fogn', { read_groups => { grp1 => 1, grp3 => 1 }},
['-R', $fogn], 0],
['rg_both', { read_groups => { grp1 => 1, grp2 => 1, grp3 => 1 }},
['-R', $fogn, '-r', 'grp2'], 0],
['rg_both2', { read_groups => { grp1 => 1, grp2 => 1, grp3 => 1 }},
['-r', 'grp2', '-R', $fogn], 0],
# Read names
['rn', { read_names => { 'unaligned_grp3_p001' => 1, 'ref1_grp1_p001' => 1, 'r008' => 1, 'r009' => 1 } },
['-N', $forn], 0],
# Tag with values
['tv_BC', { tag => 'BC', tag_values => { ACGT => 1, TGCA => 1, AATTCCGG => 1 }},
['-d', 'BC'], 0],
['tv_BC_TGCA', { tag => 'BC', tag_values => { TGCA => 1 }},
['-d', 'BC:TGCA'], 0],
['tv_BC_fobc', { tag => 'BC', tag_values => { ACGT => 1, AATTCCGG => 1 }},
['-D', "BC:${fobc}"], 0],
['tv_D_and_d', { tag => 'BC', tag_values => { ACGT => 1, TGCA => 1, AATTCCGG => 1 }},
['-D', "BC:${fobc}", '-d', 'BC:TGCA'], 0],
['tv_d_and_D', { tag => "BC", tag_values => { ACGT => 1, TGCA => 1, AATTCCGG => 1 }},
['-d', 'BC:TGCA', '-D', "BC:${fobc}"], 0],
['tv_D_RG_fogn', { tag => 'RG', tag_values => { grp1 => 1, grp3 => 1 }},
['-D', "RG:${fogn}"], 0],
['tv_d_non_existent_tag', { tag => 'NE', tag_values => { TGCA => 1 }},
['-d', 'NE:TGCA'], 0],
['tv_d_no_tag', { tag => '', tag_values => { TGCA => 1 }},
['-d', 'TGCA'], 1],
['tv_D_invalid_tag_fobc', { tag => 'BClong', tag_values => { ACGT => 1, AATTCCGG => 1 }},
['-D', "BClong:${fobc}"], 1],
['tv_d_different_tags', { tag => 'BC', tag_values => { ACGT => 1, grp2 => 1 }},
['-d', 'BC:ACGT', '-d', 'RG:grp2' ], 1],
['tv_NM_13', { tag => 'NM', tag_values => { 13 => 1 }},
['-d', 'NM:13'], 0],
['tv_ab_z', { tag => 'ab', tag_values => { z => 2 }},
['-d', 'ab:z'], 0],
# Libraries
['lib2', { libraries => { 'Library 2' => 1 }}, ['-l', 'Library 2'], 0],
['lib3', { libraries => { 'Library 3' => 1 }}, ['-l', 'Library 3'], 0],
# Mapping qualities
['mq50', { min_map_qual => 50 }, ['-q', 50], 0],
['mq99', { min_map_qual => 99 }, ['-q', 99], 0],
['mq100', { min_map_qual => 100 }, ['-q', 100], 0],
# Tag stripping
['tags1', { strip_tags => { fa => 1 } }, ['-x', 'fa'], 0],
['tags2', { strip_tags => { fa => 1, ha => 1 } },
['-x', 'fa', '-x', 'ha'], 0],
['tags2', { strip_tags => { fa => 1, ha => 1 } },
['-x', 'fa,ha'], 0],
['tags2', { strip_tags => { fa => 1, ha => 1 } },
# All tags in test file bar fa and ha, negated
['-x', '^RG,BC,NM,MD,H0,aa,ab,za,ba,bb,bc,bd,be,bf,bg,ia'], 0],
['tags2', { strip_tags => { fa => 1, ha => 1 } },
['--keep-tag', 'RG,BC,NM,MD,H0,aa,ab,za,ba,bb,bc,bd,be,bf,bg,ia'], 0],
# Tag strip plus read group
['tags_rg1', { strip_tags => { fa => 1 }, read_groups => { grp2 => 1 }},
['-x', 'fa', '-r', 'grp2'], 0],
['tags_rg2', { strip_tags => { RG => 1 }, read_groups => { grp2 => 1 }},
['-x', 'RG', '-r', 'grp2'], 0],
# Minimum query length
['qlen10', { min_qlen => 10 }, ['-m', 10], 0],
['qlen11', { min_qlen => 11 }, ['-m', 11], 0],
['qlen15', { min_qlen => 15 }, ['-m', 15], 0],
['qlen16', { min_qlen => 16 }, ['-m', 16], 0],
# Filter expressions
['expr_rej128req2', { flags_rejected => 128, flags_required => 2 },
['-e', '!(flag & 128) && (flag & 2)'], 0],
# filter_sam also removes the header line, so cannot compare.
# ['expr_RG', { read_groups => {grp1 => 1, grp3 => 1}}, ['-e', '[RG]=~"^grp[13]$"'], 0],
['expr_BC', { tag => 'BC', tag_values => { ACGT => 1, TGCA => 1, AATTCCGG => 1 }},
['-e', '[BC]'], 0],
['expr_BC2', { tag => 'BC', tag_values => { ACGT => 1, AATTCCGG => 1 }},
['-e', '[BC] == "ACGT" || [BC] == "AATTCCGG"'], 0],
['expr_mq50', { min_map_qual => 50 }, ['-e', 'mapq >= 50' ], 0],
['expr_mq99', { min_map_qual => 99 }, ['-e', 'mapq >= 99' ], 0],
['expr_mq100', { min_map_qual => 100 }, ['-e', 'mapq >= 100'], 0],
# TODO: add library to filter expression? It needs to go via RG.
# TODO: add cigar.qbase and cigar.rbase counts for consumes
# N bases of query and ref? Not the same as qlen/rlen as
# indels don't count the same.
);
my @filter_inputs = ([SAM => $sam_with_ur],
[BAM => $bam_with_ur_out],
[CRAM => $cram_with_ur_out]);
foreach my $filter (@filter_tests) {
my $sam_file = "$$opts{tmp}/view.001.$$filter[0].sam";
my ($total, $accepted) = filter_sam($sam_with_ur, $sam_file,
$$filter[1]);
foreach my $ip (@filter_inputs) {
# Filter test
my $expect_fail = $$filter[3];
my $count_output = sprintf("%s.test%03d.counts.json", $out, $test);
my $save_counts_args = ($expect_fail
? [] : ['--save-counts', $count_output]);
my $save_counts_params = ($expect_fail ? {} : {
check_save_counts => [$count_output, $total, $accepted ]
});
run_view_test($opts,
msg => "$test: Filter @{$$filter[2]} ($$ip[0] input)",
args => ['-h', @{$$filter[2]},
@$save_counts_args,
$$ip[1], '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_file,
expect_fail => $expect_fail,
%$save_counts_params);
$test++;
# Count test
run_view_test($opts,
msg => "$test: Count @{$$filter[2]} ($$ip[0] input)",
args => ['-c', @{$$filter[2]}, $$ip[1], '--no-PG'],
out => sprintf("%s.test%03d.count", $out, $test),
redirect => 1,
compare_count => $sam_file,
expect_fail => $expect_fail);
$test++;
}
}
# Region query tests
my $sam_no_ur2 = "$$opts{path}/dat/view.002.sam";
my $sam_with_ur2 = "$$opts{tmp}/view.002.sam";
add_ur_tags($sam_no_ur2, $sam_with_ur2, "$$opts{path}/dat/view.002.fa");
my $bam_with_ur_out2 = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: 1bp reads file SAM -> BAM -> SAM",
args => ['-b', $sam_with_ur2, '--no-PG'],
out => $bam_with_ur_out2,
compare_sam => $sam_with_ur2);
$test++;
my $cram_with_ur_out2 = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: 1bp reads file SAM -> CRAM -> SAM",
args => ['-C', $sam_with_ur2, '--no-PG'],
out => $cram_with_ur_out2,
compare_sam => $sam_with_ur2);
my @region_sams = ($sam_with_ur2, $sam_with_ur);
my @region_inputs = ([BAM => [$bam_with_ur_out2, $bam_with_ur_out]],
[CRAM => [$cram_with_ur_out2, $cram_with_ur_out]]);
# Add indicies
cmd("'$$opts{bin}/samtools' index '$bam_with_ur_out'");
cmd("'$$opts{bin}/samtools' index '$cram_with_ur_out'");
cmd("'$$opts{bin}/samtools' index '$bam_with_ur_out2'");
cmd("'$$opts{bin}/samtools' index '$cram_with_ur_out2'");
my $bed1 = "$$opts{path}/dat/view.001.01.bed";
my $bed2 = "$$opts{path}/dat/view.001.02.bed";
my $bed3 = "$$opts{path}/dat/view.002.01.bed";
my $bed4 = "$$opts{path}/dat/view.002.02.bed";
my $bed5 = "$$opts{path}/dat/view.001.03.bed"; # unsorted
my $bed1reg = [['ref1', 11, 24], ['ref1', 45, 45], ['ref2', 17, 17]];
my @region_tests = (
['1bp_reg1', 0, { region => [['Z']]}, [], ['Z']],
['1bp_reg2', 0, { region => [['Z', 30]]}, [], ['Z:30']],
['1bp_reg3', 0, { region => [['Z', 20, 40]]}, [], ['Z:20-40']],
['reg1', 1, { region => [['ref1']] }, [], ['ref1']],
['reg2', 1, { region => [['ref1', 15]] }, [], ['ref1:15']],
['reg3', 1, { region => [['ref1', 15, 45]] }, [], ['ref1:15-45']],
['reg4', 1, { region => [['ref1', 15, 45], ['ref2']] },
[], ['ref1:15-45', 'ref2']],
['reg5', 1, { region => [['ref1', 15, 45], ['ref2', 16]] },
[], ['ref1:15-45', 'ref2:16']],
['reg6', 1, { region => [['ref1', 15, 45], ['ref2', 16, 31]] },
[], ['ref1:15-45', 'ref2:16-31']],
['reg7', 1, { region => [['ref1', 15, 15], ['ref1', 45, 45]] },
[], ['ref1:15-15', 'ref1:45-45']],
# Regions combined with other filters.
['reg8', 1, { region => [['ref1']], flags_required => 128 },
['-f', 128], ['ref1']],
['reg9', 1, { region => [['ref1', 15, 45]], min_map_qual => 50 },
['-q', 50], ['ref1:15-45']],
['reg10', 1,
{ region => [['ref1', 15, 45]], read_groups => { grp2 => 1 }},
['-r', 'grp2'], ['ref1:15-45']],
# Unmapped reads
['reg_unmapped1', 1, { region => [['*']] }, [], ['*']],
# Regions from BED files. Regions here need to be kept in synch.
# with the .bed files in test/dat. Note that BED counts from
# base 0 and ends ranges one past the last base. Note also that
# samtools also understands a two-column format that describes a
# single point, and in that case the position is 1-based. And
# even better, you can mix the two in the same file. Confusing? Yes.
#
# $bed1reg is defined above as it's used a lot of times.
['1bp_bed3', 0, { region => [['Z', 20, 40]]}, ['-L', $bed3], []],
['1bp_bed4', 0,
{ region => [['Z', 10, 15], ['Z', 20, 20], ['Z', 25, 30],
['Z', 35, 35], ['Z', 40, 50]]}, ['-L', $bed4], []],
['bed1', 1, { region => $bed1reg }, ['-L', $bed1], []],
['bed2', 1, { region => [['ref1', 6, 20]] }, ['-L', $bed2], []],
['bed5', 1, { region => [['ref1', 11, 24], ['ref1', 45, 45],
['ref2', 12, 12], ['ref2', 47, 47]]},
['-L', $bed5], []],
# BED file plus region specification.
['1bp_bed3r1', 0, { region => [['Z', 20, 40]]},
['-L', $bed3], ['Z']],
['1bp_bed3r2', 0, { region => [['Z', 30, 40]]},
['-L', $bed3], ['Z:30']],
['1bp_bed3r3', 0, { region => [['Z', 20, 30]]},
['-L', $bed3], ['Z:10-30']],
['1bp_bed3r4', 0, { region => [['Z', 25, 35]]},
['-L', $bed3], ['Z:25-35']],
['bed1r1', 1, { region => [['ref1', 11, 16]] },
['-L', $bed1], ['ref1:11-16']],
['bed1r2', 1,
{ region => [['ref2', 17, 17]] }, ['-L', $bed1], ['ref2']],
['bed1r11', 1,
{ region => [['ref1', 11, 15], ['ref1', 45, 45]]}, ['-L', $bed1],
['ref1:1-15', 'ref1:44-45']],
# BED file plus other filters
['bed1f1', 1, { region => $bed1reg, read_groups => { grp1 => 1} },
['-L', $bed1, '-r', 'grp1'], []],
['bed1f2', 1, { region => $bed1reg, flags_required => 128 },
['-L', $bed1, '-f', 128], []],
['bed1f3', 1, { region => $bed1reg, min_map_qual => 5 },
['-L', $bed1, '-q', 5], []],
# BED file, region and filters
['bed1f1', 1,
{ region => [['ref1', 11, 16]], read_groups => { grp1 => 1}},
['-L', $bed1, '-r', 'grp1'], ['ref1:11-16']],
# BED file with multi-region iterator. The result should be the
# same as with the normal '-L' option. The '--unmap' option is
# included to ensure that the index jumping is working (if not
# the output will include more reads than expected).
['bed1M1', 1, { region => $bed1reg },
['-L', $bed1, '-M', '--unmap'], []],
# Intersection of a BED file with a region.
['bed1M2', 1, { region => [['ref1', 11, 24]] },
['-L', $bed1, '-M', '--unmap'], ['ref1:11-30']],
);
foreach my $rt (@region_tests) {
my $input_sam = $region_sams[$$rt[1]];
my $sam_file = "$$opts{tmp}/view.$$rt[0].sam";
filter_sam($input_sam, $sam_file, $$rt[2]);
foreach my $ip (@region_inputs) {
my $input_file = $$ip[1]->[$$rt[1]];
# Region test
run_view_test($opts,
msg => "$test: Region @{$$rt[3]} @{$$rt[4]} ($$ip[0] input)",
args => ['-h', @{$$rt[3]}, $input_file, @{$$rt[4]}, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_file);
$test++;
# Count test
run_view_test($opts,
msg => "$test: Count @{$$rt[3]} @{$$rt[4]} ($$ip[0] input)",
args => ['-c', @{$$rt[3]}, $input_file, @{$$rt[4]}, '--no-PG'],
out => sprintf("%s.test%03d.count", $out, $test),
redirect => 1,
compare_count => $sam_file);
$test++;
}
}
# -L option with nested regions
run_view_test($opts,
msg => "$test: -L with nested regions",
args => ['-h', '-L', "$$opts{path}/dat/nested.bed", '--no-PG',
"$$opts{path}/dat/large_chrom.sam"],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => "$$opts{path}/dat/nested.expected.sam");
$test++;
# -T / -t options
my $sam_no_sq = "$$opts{tmp}/view.001.no_sq.sam";
filter_sam($sam_no_ur, $sam_no_sq, {no_sq => 1});
my $sam_no_m5 = "$$opts{tmp}/view.001.no_m5.sam";
filter_sam($sam_no_ur, $sam_no_m5, {no_m5 => 1});
# We can't make a BAM without @SQ lines. Instead make one
# with no M5/UR in @SQ and see if we can still use it to make CRAM.
my $bam_no_m5 = "$$opts{tmp}/view.001.no_sq.bam";
run_view_test($opts,
msg => "$test: Make BAM with no M5/UR tags",
args => ['-b', $sam_no_m5, '--no-PG'],
out => $bam_no_m5,
compare_sam => $sam_no_m5);
$test++;
my $ref_file = "$$opts{path}/dat/view.001.fa";
my $ref_idx = "$$opts{path}/dat/view.001.fa.fai";
# Test SAM output
foreach my $in ([SAM => $sam_no_sq], [BAM => $bam_no_m5]) {
foreach my $topt (['-t', $ref_idx], ['-T', $ref_file]) {
run_view_test($opts,
msg => "$test: Add \@SQ with $topt->[0] ($in->[0] -> SAM)",
args => ['-h', @$topt, $in->[1], '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_no_m5);
$test++;
}
}
# Test BAM output.
foreach my $in ([SAM => $sam_no_sq], [BAM => $bam_no_m5]) {
foreach my $topt (['-t', $ref_idx], ['-T', $ref_file]) {
my $bam = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: Add \@SQ with $topt->[0] ($in->[0] -> BAM)",
args => ['-b', @$topt, $in->[1], '--no-PG'],
out => $bam,
compare_sam => $sam_no_m5);
$test++;
}
}
# Test CRAM output
foreach my $in ([SAM => $sam_no_sq], [BAM => $bam_no_m5]) {
foreach my $topt (['-t', $ref_idx], ['-T', $ref_file]) {
my $cram = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: Add \@SQ with $topt->[0] ($in->[0] -> CRAM)",
args => ['-C', @$topt, $in->[1], '--no-PG'],
out => $cram,
compare_sam => $sam_with_ur);
$test++;
}
}
# Test CRAM with explicit -T
my $cram_no_ur_t = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: Make CRAM with no UR field",
args => ['-C', $sam_no_ur, '--no-PG'],
ref_path => "$$opts{path}/dat/cram_md5",
out => $cram_no_ur_t);
$test++;
run_view_test($opts,
msg => "$test: Decoding CRAM with no UR field via -T",
args => ['-T', $ref_file, $cram_no_ur_t, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $sam_no_ur);
$test++;
# CIGAR B-operator removal tests.
my $b_op_sam = "$$opts{path}/dat/view.003.sam";
my $b_op_expected = "$$opts{path}/dat/view.003.expected.sam";
run_view_test($opts,
msg => "$test: CIGAR B-operator removal",
args => ['-h', '-B', $b_op_sam, '--no-PG'],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $b_op_expected);
$test++;
# Threads
# big SAM -> BAM
my $big_bam = sprintf("%s.test%03d.bam", $out, $test);
run_view_test($opts,
msg => "$test: Big SAM -> BAM",
args => ['-b', $big_sam, '--no-PG'],
out => $big_bam);
$test++;
foreach my $threads (2, 4) {
run_view_test($opts,
msg => "$test: Big SAM -> BAM ($threads threads)",
args => ['-b', '-@', $threads, $big_sam, '--no-PG'],
out => sprintf("%s.test%03d.bam", $out, $test),
compare_bam => $big_bam);
$test++;
}
# big SAM -> CRAM
my $big_cram = sprintf("%s.test%03d.cram", $out, $test);
run_view_test($opts,
msg => "$test: Big SAM -> CRAM",
args => ['-C', $big_sam, '--no-PG'],
out => $big_cram,
compare_sam => $big_sam,
pipe => 1);
$test++;
foreach my $threads (2, 4) {
run_view_test($opts,
msg => "$test: Big SAM -> CRAM ($threads threads)",
args => ['-C', '-@', $threads, $big_sam, '--no-PG'],
out => sprintf("%s.test%03d.cram", $out, $test),
compare_sam => $big_sam,
pipe => 1);
$test++;
}
# Subsampling (-s) option. This is random so accept if within +/- 5%
# of the expected number of reads.
foreach my $ip ([SAM => $big_sam], [BAM => $big_bam], [CRAM => $big_cram]) {
foreach my $frac (0.2, 0.5, 0.8) {
run_view_subsample_test($opts,
msg => "$test: Subsample $frac ($ip->[0] input)",
frac => $frac,
input => $ip->[1],
trials => 10,
min => $big_sam_count * $frac * 0.95,
max => $big_sam_count * $frac * 1.05);
$test++;
}
}
my $b_pg_sam = "$$opts{path}/dat/view.001.sam";
my $b_pg_expected = "$$opts{path}/dat/view.004.expected.sam";
run_view_test($opts,
msg => "$test: SAM add PG",
args => ['-h', $b_pg_sam],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $b_pg_expected);
# unset flags and clear tags associated with duplication
$test++;
my $dup_sam = "$$opts{path}/dat/view.005.sam";
my $dup_expected = "$$opts{path}/dat/view.005.expected.sam";
run_view_test($opts,
msg=> "$test: Unset dup flag, remove dt and do tags",
args => ['-h', '--remove-flags', 'DUP', '-x', 'do', '-x', 'dt', '--no-PG', $dup_sam],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $dup_expected);
# unmap excluded reads, ones marked as duplicate in this case
$test++;
my $unmapped_expected = "$$opts{path}/dat/view.005.unmap.expected.sam";
run_view_test($opts,
msg=> "$test: Unmap dup flagged reads.",
args => ['-h', '-F', 'DUP', '-p', '--no-PG', $dup_sam],
out => sprintf("%s.test%03d.sam", $out, $test),
compare => $unmapped_expected);
$test++;
run_view_test($opts,
msg=> "$test: Unmap dup flagged reads.",
args => ['-c', '-F', 'DUP', '-p', '--no-PG', $dup_sam],
out => sprintf("%s.test%03d.sam", $out, $test),
compare_count => 2);
# retrieve reads from a region including their mates
my $count_output;
my $bam = 'test/dat/view.fetch-pairs.bam';
cmd("$$opts{bin}/samtools index $bam");
$test++;
$count_output = sprintf("%s.fetch-pairs.test%03d.counts.json", $out, $test);
run_view_test($opts,
msg => "$test: fetch pairs",
args => ['--no-PG', '--save-counts', $count_output,
'--fetch-pairs',$bam,'6:25515943-25515943','6:25020026-25020026','6:25515822-25515822'],
out => sprintf("%s.fetch-pairs.test%03d.bam", $out, $test),
compare_sam => 'test/dat/view.fetch-pairs.expected.sam',
check_save_counts => [$count_output, 28, 28]);
$test++;
$count_output = sprintf("%s.fetch-pairs.test%03d.counts.json", $out, $test);
run_view_test($opts,
msg => "$test: fetch pairs",
args => ['--no-PG', '--save-counts', $count_output,
'--fetch-pairs',$bam,'6:25515857-25515857'],
out => sprintf("%s.fetch-pairs.test%03d.bam", $out, $test),
compare_sam => 'test/dat/view.fetch-pairs.filter0.expected.sam',
check_save_counts => [$count_output, 34, 34]);
$test++;
$count_output = sprintf("%s.fetch-pairs.test%03d.counts.json", $out, $test);
run_view_test($opts,
msg => "$test: fetch pairs",
args => ['--no-PG', '--save-counts', $count_output,
'--fetch-pairs','--exclude-flags','DUP',$bam,'6:25515857-25515857'],
out => sprintf("%s.fetch-pairs.test%03d.bam", $out, $test),
compare_sam => 'test/dat/view.fetch-pairs.filter1.expected.sam',
check_save_counts => [$count_output, 31, 28]);
}
sub gen_head_output
{
my ($opts, $h, $n, $desc, $infile) = @_;
open my $in, '<', $infile or die "Couldn't open $infile: $!\n";
my $expected = "dat/head.$desc.tmp.expected";
open my $out, '>', "$$opts{path}/$expected"
or die "Couldn't write to $expected: $!\n";
while (<$in>) {
my $counter = /^@/? \$h : \$n;
next unless $$counter > 0;
print $out $_;
$$counter--;
}
close $in;
close $out;
return $expected;
}
sub test_head
{
my ($opts) = @_;
my $infile = "$$opts{path}/dat/view.001.sam";
test_cmd($opts, out => gen_head_output($opts, 1000, 0, "all", $infile),
cmd => "$$opts{bin}/samtools head $infile");
test_cmd($opts, out => gen_head_output($opts, 0, 0, "none", $infile),
cmd => "$$opts{bin}/samtools head -h 0 $infile");
test_cmd($opts, out => gen_head_output($opts, 1, 0, "one", $infile),
cmd => "$$opts{bin}/samtools head -h 1 $infile");
test_cmd($opts, out => gen_head_output($opts, 5, 0, "five", $infile),
cmd => "$$opts{bin}/samtools head -h 5 $infile");
# view.001.sam has 29 header lines; test exactly that and asking for 1 extra
test_cmd($opts, out => gen_head_output($opts, 29, 0, "exact", $infile),
cmd => "$$opts{bin}/samtools head -h 29 $infile");
test_cmd($opts, out => gen_head_output($opts, 30, 0, "toomany", $infile),
cmd => "$$opts{bin}/samtools head -h 30 $infile");
test_cmd($opts, out => gen_head_output($opts, 1000, 0, "alln0", $infile),
cmd => "$$opts{bin}/samtools head -n 0 $infile");
test_cmd($opts, out => gen_head_output($opts, 1000, 1, "onerec", $infile),
cmd => "$$opts{bin}/samtools head -n 1 $infile");
test_cmd($opts, out => gen_head_output($opts, 1000, 5, "fiverecs", $infile),
cmd => "$$opts{bin}/samtools head -n 5 $infile");
test_cmd($opts, out => gen_head_output($opts, 5, 5, "fiveboth", $infile),
cmd => "$$opts{bin}/samtools head -h 5 -n 5 < $infile");
}
# cat SAM files in the same way as samtools cat does with BAMs
#
# $sam_out is the name of the output file
# $sam_in is the name of the input file. More than one of these can be
# passed in. The header is taken from the first file.
sub cat_sams
{
my $sam_out = shift(@_);
my $first = 1;
open(my $out, '>', $sam_out)
|| die "Couldn't open $sam_out for writing: $!\n";
while (my $sam_in = shift(@_)) {
my $in;
if ($sam_in =~ /\.gz$/) {
$in = open_bgunzip($opts, $sam_in);
} else {
open($in, '<', $sam_in) || die "Couldn't open $sam_in : $!\n";
}
if ($first) {
# copy header
my $prev = '';
while (<$in>) {
if (/^@/) { print $out $_ || die "Error writing to $sam_out : $!\n"; }
else { $prev = $_; last; }
}
print $out "\@PG\tID:samtools\tPN:samtools\n";
if ($prev) { print $out $prev; }
}
while (<$in>) {
next if (/^@/ && !$first);
print $out $_ || die "Error writing to $sam_out : $!\n";
}
close($in) || die "Error reading $sam_in : $!\n";
$first = 0;
}
close($out) || die "Error writing to $sam_out : $!\n";
}
# Large position tests
sub test_large_positions
{
my ($opts) = @_;
# Ensure the tview test prints out the expected number of columns
local $ENV{COLUMNS} = 80;
# Simple round-trip
my $longref = "$$opts{tmp}/longref.sam.gz";
cmd("$$opts{bgzip} -c $$opts{path}/large_pos/longref.sam > $longref");
test_cmd($opts, out => 'large_pos/longref.sam',
cmd => "$$opts{bin}/samtools view -h --no-PG $longref");
# Index
cmd("$$opts{bin}/samtools index -c $longref");
test_cmd($opts, out => 'large_pos/longref_idx.expected.sam',
cmd => "$$opts{bin}/samtools view -h --no-PG $longref CHROMOSOME_I:10000000114-10000000168");
# Bed file
test_cmd($opts, out => 'large_pos/longref_idx.expected.sam',
cmd => "$$opts{bin}/samtools view -h --no-PG -L $$opts{path}/large_pos/test.bed $longref");
# Sort
test_cmd($opts, out => 'large_pos/longref.sam',
cmd => "$$opts{bin}/samtools sort -O sam --no-PG -m 10M $$opts{path}/large_pos/longref_name.sam");
# Merge
test_cmd($opts, out => 'large_pos/merge.expected.sam',
cmd => "$$opts{bin}/samtools merge -O sam --no-PG - $$opts{path}/large_pos/longref.sam $$opts{path}/large_pos/longref2.sam");
# Depth
test_cmd($opts, out => 'large_pos/depth.expected.out',
cmd => "$$opts{bin}/samtools depth $$opts{path}/large_pos/longref.sam");
test_cmd($opts, out => 'large_pos/depth_bed.expected.out',
cmd => "$$opts{bin}/samtools depth -b $$opts{path}/large_pos/test.bed $$opts{path}/large_pos/longref.sam");
# tview
test_cmd($opts, out => 'large_pos/tview.expected.out',
cmd => "$$opts{bin}/samtools tview -d T -p CHROMOSOME_I:10000000000 $longref");
# Sort and fixmates
test_cmd($opts, out => 'large_pos/longref3.expected.sam',
cmd => "$$opts{bin}/samtools sort -O sam --no-PG -n -m 10M test/large_pos/longref3.sam |
$$opts{bin}/samtools fixmate -O sam --no-PG - - |
$$opts{bin}/samtools sort -O sam --no-PG -m 10M");
}
# Test samtools cat.
sub test_cat
{
my ($opts) = @_;
my $test_name = "test_cat";
print "$test_name:\n";
my @sams;
my @sams_r;
my @bams;
my @crams;
my @bgbams;
my $nfiles = 4;
my $test = 1;
my $out = "$$opts{tmp}/cat";
# Generate some files big enough to include a few bgzf blocks
for (my $i = 0; $i < $nfiles; $i++) {
($sams[$i]) = gen_file($opts, sprintf("%s.%d", $out, $i + 1), 10000, 15551);
# Convert to BAM
$bams[$i] = sprintf("%s.%d.bam", $out, $i + 1);
$sams_r[$i] = sprintf("%s.%d.reg.sam", $out, $i + 1);
run_view_test($opts,
msg => sprintf("Generate BAM file #%d", $i + 1),
args => ['-b', $sams[$i], '--no-PG', '--write-index'],
out => $bams[$i],
compare_sam => $sams[$i],
pipe => 1);
run_view_test($opts,
msg => sprintf("Generate BAM region file #%d", $i + 1),
args => ['-h', $bams[$i], '--no-PG', 'ref1:4240-7150'],
out => "$sams_r[$i]",
pipe => 1);
# Recompress with bgzip to alter the location of the bgzf boundaries.
$bgbams[$i] = sprintf("%s.%d.bgzip.bam", $out, $i + 1);
cmd("'$$opts{bgzip}' -c -d < '$bams[$i]' | '$$opts{bgzip}' -c > '$bgbams[$i]'");
# Create CRAMs
$crams[$i] = sprintf("%s.%d.cram", $out, $i + 1);
run_view_test($opts,
msg => sprintf("Generate CRAM file #%d", $i + 1),
args => ['-O', 'CRAM,seqs_per_slice=100', $sams[$i],
'--no-PG', '--write-index'],
out => $crams[$i],
compare_sam => $sams[$i],
pipe => 1);
}
# Make a concatenated SAM file to compare
my $catsam1 = "$out.all1.sam";
my $catsam1r = "$out.all1r.sam";
cat_sams($catsam1, @sams);
cat_sams($catsam1r, @sams_r);
foreach my $redirect (0, 1) {
my $to_stdout = $redirect ? ' to stdout' : '';
# Basic test
run_view_test($opts,
msg => "$test: cat BAM files$to_stdout",
cmd => 'cat',
args => [@bams],
out => sprintf("%s.test%03d.bam", $out, $test),
redirect => $redirect,
compare_sam => $catsam1);
$test++;
# Test BAM files recompressed with bgzip
run_view_test($opts,
msg => "$test: cat recompressed BAM files$to_stdout",
cmd => 'cat',
args => [@bgbams],
out => sprintf("%s.test%03d.bam", $out, $test),
redirect => $redirect,
compare_sam => $catsam1);
$test++;
# Test CRAM files
run_view_test($opts,
msg => "$test: cat CRAM files$to_stdout",
cmd => 'cat',
args => [@crams],
out => sprintf("%s.test%03d.cram", $out, $test),
redirect => $redirect,
compare_sam => $catsam1);
$test++;
# Test CRAM sub-regions
run_view_test($opts,
msg => "$test: cat CRAM subregion files$to_stdout",
cmd => 'cat',
args => ['-r', 'ref1:4240-7150', @crams],
out => sprintf("%s.test%03d.cram", $out, $test),
redirect => $redirect,
compare_sam => $catsam1r);
$test++;
}
# Test CRAM -p option. Do 1/2 and 2/2 and check the combined result matches
run_view_test($opts,
msg => "$test: cat CRAM part 1/2",
cmd => 'cat',
args => ['-p', '1/2', @crams],
out => sprintf("%s.test_p1.cram", $out));
run_view_test($opts,
msg => "$test: cat CRAM part 2/2",
cmd => 'cat',
args => ['-p', '2/2', @crams],
out => sprintf("%s.test_p2.cram", $out));
run_view_test($opts,
msg => "$test: cat CRAM parts 1/2 + 2/2",
cmd => 'cat',
args => ['--no-PG',
sprintf("%s.test_p1.cram", $out),
sprintf("%s.test_p2.cram", $out)],
out => sprintf("%s.test_p12.cram", $out),
compare_sam => $catsam1);
$test++;
# Test reheader option
my $hdr_no_ur = "$$opts{path}/dat/cat.hdr";
my $header = "$$opts{tmp}/cat.hdr";
add_ur_tags($hdr_no_ur, $header, "$$opts{tmp}/cat.1.fa");
my $catsam2 = "$out.all2.sam";
cat_sams($catsam2, $header, @sams);
run_view_test($opts,
msg => "$test: cat BAM files with new header",
cmd => 'cat',
args => ['-h', $header, @bams],
out => sprintf("%s.test%03d.bam", $out, $test),
compare_sam => $catsam2);
$test++;
run_view_test($opts,
msg => "$test: cat recompressed BAM files with new header",
cmd => 'cat',
args => ['-h', $header, @bgbams],
out => sprintf("%s.test%03d.bam", $out, $test),
compare_sam => $catsam2);
$test++;
run_view_test($opts,
msg => "$test: cat CRAM files with new header",
cmd => 'cat',
args => ['-h', $header, @crams],
out => sprintf("%s.test%03d.cram", $out, $test),
compare_sam => $catsam2);
$test++;
}
sub sam2fq
{
my ($sam_in, $fq_out, $suffixes) = @_;
open(my $in, '<', $sam_in) || die "Couldn't open $sam_in : $!\n";
open(my $out, '>', $fq_out)
|| die "Couldn't open $fq_out for writing : $!\n";
while (<$in>) {
next if (/^@/);
my @s = split(/\t/, $_);
next if ($s[1] & (256|2048));
my $dirn = ($s[1] & 0xc0) >> 6;
my $suff = $suffixes ? ('', '/1', '/2', '')[$dirn] : '';
if (($s[1] & 0x10) != 0) { # reverse complement
$s[9] =~ tr/ACGTMRWSYKVHDBN/TGCAKYWSRMBDHVN/;
$s[9] = reverse($s[9]);
$s[10] = reverse($s[10]);
}
print $out "\@$s[0]$suff\n$s[9]\n+\n$s[10]\n";
}
close($out) || die "Error writing $fq_out : $!\n";
close($in) || die "Error reading $sam_in : $!\n";
}
# Conversion of FASTQ to BAM.
# We use the bam2fq expected output to validate on.
# This permits round trip validation
sub test_import
{
my ($opts, %args) = @_;
# Just 1 end, as an unpaired read sample; eg as if ont or pacbio.
# -0 or implicit (lack of /1 /2 suffixes) via -s.
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"0.fq" => 'bam2fq/1.1.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -0 test/bam2fq/1.1.fq.expected | $$opts{bin}/samtools fastq -0 $$opts{path}/0.fq");
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"0.fq" => 'bam2fq/1.1.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -s test/bam2fq/1.1.fq.expected | $$opts{bin}/samtools fastq -0 $$opts{path}/0.fq");
# Just 1 end, as half of a paired-end sample. Can be either explicit via
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"s.fq" => 'bam2fq/5.s.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -s test/bam2fq/5.s.fq.expected | $$opts{bin}/samtools fastq -s $$opts{path}/s.fq");
# Normal read 1 / read 2
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/1.1.fq.expected',
"2.fq" => 'bam2fq/1.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG test/bam2fq/1.1.fq.expected test/bam2fq/1.2.fq.expected | $$opts{bin}/samtools fastq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
# Normal read 1 / read 2 but with /1 and /2 suffixes.
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/5.1.fq.expected',
"2.fq" => 'bam2fq/5.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG test/bam2fq/5.1.fq.expected test/bam2fq/5.2.fq.expected | $$opts{bin}/samtools fastq -N -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
# Barcodes via CASAVA tags
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/12.1.fq.expected',
"2.fq" => 'bam2fq/12.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -i -1 test/bam2fq/12.1.fq.expected -2 test/bam2fq/12.2.fq.expected | $$opts{bin}/samtools fastq -i --index-format 'i*i*' -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/12.1.fq.expected',
"2.fq" => 'bam2fq/12.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG --barcode-tag OX -i -1 test/bam2fq/12.1.fq.expected -2 test/bam2fq/12.2.fq.expected | $$opts{bin}/samtools fastq --barcode-tag OX -i --index-format 'i*i*' -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
# Barcodes via explicit aux tags; 6
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/6.1.fq.expected',
"2.fq" => 'bam2fq/6.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -T \"\" -1 test/bam2fq/6.1.fq.expected -2 test/bam2fq/6.2.fq.expected | $$opts{bin}/samtools fastq -N -T RG,BC,QT -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
# Other aux tags; 7
test_cmd($opts, out=>'dat/empty.expected',
out_map=>{"1.fq" => 'bam2fq/7.1.fq.expected',
"2.fq" => 'bam2fq/7.2.fq.expected'},
cmd=>"$$opts{bin}/samtools import --no-PG -T \"*\" -1 test/bam2fq/7.1.fq.expected -2 test/bam2fq/7.2.fq.expected | $$opts{bin}/samtools fastq -N -T RG,BC,QT,MD,ia -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq");
#------------------------
# Plus our own test files, using bam2fq as source
# Read-group
test_cmd($opts, out=>'import/1.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/bam2fq/1.1.fq.expected test/bam2fq/1.2.fq.expected -R rgid");
test_cmd($opts, out=>'import/1.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/bam2fq/1.1.fq.expected test/bam2fq/1.2.fq.expected -r ID:rgid");
test_cmd($opts, out=>'import/1.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/bam2fq/1.1.fq.expected test/bam2fq/1.2.fq.expected -r '\@RG\tID:rgid'");
# Interleaved data
test_cmd($opts, out=>'import/2.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/import/2.interleaved.fq -T \"\"");
test_cmd($opts, out=>'import/2.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/import/3.interleaved.fq -i");
# Non aux-tag comments (we don't use these, but also shouldn't choke).
test_cmd($opts, out=>'import/4.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/import/4.aux.fq -T \"*\"");
test_cmd($opts, out=>'import/4.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/import/4.aux.fq -T \"\"");
test_cmd($opts, out=>'import/4.expected-XZ,XA,AA.sam',
cmd=>"$$opts{bin}/samtools import --no-PG test/import/4.aux.fq -T XZ,XA,AA");
# Barcode files
test_cmd($opts, out=>'import/5-BC.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG --i1 test/import/5-i1.fq --i2 test/import/5-i2.fq --r1 test/import/5-r1.fq --r2 test/import/5-r2.fq");
test_cmd($opts, out=>'import/5-OX.expected.sam',
cmd=>"$$opts{bin}/samtools import --no-PG --i1 test/import/5-i1.fq --i2 test/import/5-i2.fq --r1 test/import/5-r1.fq --r2 test/import/5-r2.fq --barcode-tag OX --quality-tag BZ");
}
sub test_bam2fq
{
my ($opts, %args) = @_;
my $threads = exists($args{threads}) ? ['-@', $args{threads}] : [];
my $nthreads = exists($args{threads}) ? " ($args{threads} threads)" : "";
my $test_name = "test_bam2fq";
print "$test_name:\n";
my $out = "$$opts{tmp}/bam2fq";
my $sam = "$$opts{path}/dat/bam2fq.001.sam";
my $bam = "$out.001.bam";
my $cram = "$out.001.cram";
my $fqsuffix = "$out.001.fq";
my $fqnosuffix = "$out.001.nosuff.fq";
sam2fq($sam, $fqsuffix, 1);
sam2fq($sam, $fqnosuffix, 0);
# Make a BAM file with the test data
run_view_test($opts,
msg => "Generate BAM file$nthreads",
args => ['-b', $sam, @$threads, '--no-PG'],
out => $bam,
compare_sam => $sam,
pipe => 1);
# Make a CRAM file with the test data
run_view_test($opts,
msg => "Generate CRAM file$nthreads",
args => ['-C', $sam, @$threads, '--no-PG'],
out => $cram,
ref_path => "$$opts{path}/dat/cram_md5",
compare_sam => $sam,
pipe => 1);
my $test = 1;
my @inputs = ([SAM => $sam], [BAM => $bam], [CRAM => $cram]);
foreach my $input (@inputs) {
foreach my $nosuffix (0, 1) {
my @n = $nosuffix ? ('-n') : ();
run_view_test($opts,
msg => "$test: fastq @n ($input->[0] input)$nthreads",
cmd => 'fastq',
args => [@n, @$threads, $input->[1]],
out => sprintf("%s.test%03d.fq", $out, $test),
ref_path => "$$opts{path}/dat/cram_md5",
redirect => 1,
compare_text => $nosuffix ? $fqnosuffix : $fqsuffix);
$test++;
}
}
# basic 2 output test without singleton tracking
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/1.1.fq.expected', '2.fq' => 'bam2fq/1.2.fq.expected'},cmd=>"$$opts{bin}/samtools fastq @$threads -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.001.sam");
# basic 2 output test with singleton tracking but no singleton
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/2.1.fq.expected', '2.fq' => 'bam2fq/2.2.fq.expected', 's.fq' => 'bam2fq/2.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.001.sam");
# basic 2 output test with singleton tracking with a singleton in the middle
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/3.1.fq.expected', '2.fq' => 'bam2fq/3.2.fq.expected', 's.fq' => 'bam2fq/3.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.002.sam");
# basic 2 output test with singleton tracking with a singleton as last read
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/4.1.fq.expected', '2.fq' => 'bam2fq/4.2.fq.expected', 's.fq' => 'bam2fq/4.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.003.sam");
# tag output test with singleton tracking with a singleton as last read
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/4.1.fq.expected', '2.fq' => 'bam2fq/4.2.fq.expected', 's.fq' => 'bam2fq/4.s.fq.expected', 'bc.fq' => 'bam2fq/bc.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC --index-format 'n2i2' --i1 $$opts{path}/bc.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.004.sam");
# test -O flag with no OQ tags
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/4.1.fq.expected', '2.fq' => 'bam2fq/4.2.fq.expected', 's.fq' => 'bam2fq/4.s.fq.expected', 'bc.fq' => 'bam2fq/bc.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC -O --index-format 'n2i2' --i1 $$opts{path}/bc.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.004.sam");
# test -O flag with OQ tags
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/10.1.fq.expected', '2.fq' => 'bam2fq/10.2.fq.expected', 's.fq' => 'bam2fq/10.s.fq.expected', 'bc.fq' => 'bam2fq/bc10.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC -O --index-format 'n2i2' --i1 $$opts{path}/bc.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.010.sam");
# tag output test with separators and -N flag
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/5.1.fq.expected', '2.fq' => 'bam2fq/5.2.fq.expected', 's.fq' => 'bam2fq/5.s.fq.expected', 'bc_split.fq' => 'bam2fq/bc_split.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC -N --index-format 'n*i*' --i1 $$opts{path}/bc_split.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.005.sam");
# -t flag
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/6.1.fq.expected', '2.fq' => 'bam2fq/6.2.fq.expected', 's.fq' => 'bam2fq/6.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -N -t -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.005.sam");
# -T flag
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/7.1.fq.expected', '2.fq' => 'bam2fq/7.2.fq.expected', 's.fq' => 'bam2fq/7.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -N -t -T MD,ia -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.005.sam");
# -i flag with index
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/8.1.fq.expected', '2.fq' => 'bam2fq/8.2.fq.expected', 's.fq' => 'bam2fq/8.s.fq.expected', 'i.fq' => 'bam2fq/8.i.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC -i --index-format 'n2i2' --i1 $$opts{path}/i.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.004.sam");
# -i flag with dual index
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/12.1.fq.expected', '2.fq' => 'bam2fq/12.2.fq.expected', 's.fq' => 'bam2fq/12.s.fq.expected', 'i.fq' => 'bam2fq/12.i.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --barcode-tag BC -i --index-format 'i*i*' --i1 $$opts{path}/i.fq -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.005.sam");
# -i flag with dual index but no indexes
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/12.1.fq.expected', '2.fq' => 'bam2fq/12.2.fq.expected', 's.fq' => 'bam2fq/12.s.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -i --index-format 'i*i*' -s $$opts{path}/s.fq -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.005.sam");
# test for Issue #703 (failure to write all reads on uncollated input)
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'1.fq' => 'bam2fq/9.1.fq.expected', '2.fq' => 'bam2fq/9.2.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads -1 $$opts{path}/1.fq -2 $$opts{path}/2.fq $$opts{path}/dat/bam2fq.703.sam");
# Read 1/2 output, duplicate filename (-1 -2)
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'o.fq' => 'bam2fq/11.fq.expected'},cmd=>"$$opts{bin}/samtools fastq @$threads -N -1 $$opts{path}/o.fq -2 $$opts{path}/o.fq $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'o.fa' => 'bam2fq/11.fa.expected'},cmd=>"$$opts{bin}/samtools fasta @$threads -N -1 $$opts{path}/o.fa -2 $$opts{path}/o.fa $$opts{path}/dat/bam2fq.001.sam");
# Read 1/2 output, single filename (-o)
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'o.fq' => 'bam2fq/11.fq.expected'},cmd=>"$$opts{bin}/samtools fastq @$threads -N -o $$opts{path}/o.fq $$opts{path}/dat/bam2fq.001.sam");
# Read 1/2 output, stdout and discard singletons/other
test_cmd($opts, out=>'bam2fq/11.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -s $out.discard.s.fq -0 $out.discard.0.fq $$opts{path}/dat/bam2fq.001.sam");
# Test B aux tag
test_cmd($opts, out=>'bam2fq/13.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -T ba,bb,bc,bd,be,bf,bg $$opts{path}/dat/bam2fq.013.sam");
# Test single ended output with dual-indexing
test_cmd($opts, out=>'dat/empty.expected', out_map=>{'0.fq' => 'bam2fq/14.0.fq.expected', 'i1.fq' => 'bam2fq/14.i1.fq.expected', 'i2.fq' => 'bam2fq/14.i2.fq.expected', '0.fq' => 'bam2fq/14.0.fq.expected'}, cmd=>"$$opts{bin}/samtools fastq @$threads --index-format 'i8n1i8' --i1 $$opts{path}/i1.fq --i2 $$opts{path}/i2.fq -0 $$opts{path}/0.fq $$opts{path}/dat/bam2fq.014.sam");
# -T flag, all tags mode
test_cmd($opts, out=>'bam2fq/15.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -T '' $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'bam2fq/15.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -t -T '*' $$opts{path}/dat/bam2fq.001.sam");
# -d TAG
test_cmd($opts, out=>'bam2fq/16.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -T '*' -d MD:10 $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'bam2fq/17.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -T '*' -d NM:0 $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'bam2fq/18.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -N -T '*' -d ia $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'bam2fq/20.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -d NM:13 -d NM:14 $$opts{path}/dat/bam2fq.001.sam");
# -D TAG
test_cmd($opts, out=>'bam2fq/20.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -D NM:test/dat/bam2fq.NM-D $$opts{path}/dat/bam2fq.001.sam");
test_cmd($opts, out=>'bam2fq/19.fq.expected', cmd=>"$$opts{bin}/samtools fastq @$threads -D MD:test/dat/bam2fq.MD-D $$opts{path}/dat/bam2fq.001.sam");
}
sub test_depad
{
my ($opts) = @_;
my $test_name = "test_depad";
print "$test_name:\n";
my $pad_sam = "$$opts{path}/dat/depad.001p.sam";
my $unpad_sam = "$$opts{path}/dat/depad.001u.sam";
my $ref = "$$opts{path}/dat/depad.001.fa";
my $out = "$$opts{tmp}/depad";
my $pad_bam = "$out.001p.bam";
my $pad_cram = "$out.001p.cram";
# Make a BAM file with the test data
run_view_test($opts,
msg => "Generate BAM file",
args => ['-b', $pad_sam, '--no-PG'],
out => $pad_bam,
compare_sam => $pad_sam);
# Don't try CRAM for now as it loses the CIGAR string from the embedded
# reference.
# # Make a CRAM file with the test data
# run_view_test($opts,
# msg => "Generate CRAM file",
# args => ['-C', $pad_sam],
# out => $pad_cram,
# ref_path => "$$opts{path}/dat/cram_md5",
# compare_sam => $pad_sam);
my $test = 1;
# my @inputs = ([SAM => $pad_sam], [BAM => $pad_bam], [CRAM => $pad_cram]);
my @inputs = ([SAM => $pad_sam], [BAM => $pad_bam]);
my @formats = (['', 'bam'], ['-s', 'sam'], ['-u', 'uncomp.bam'],
['-1', 'fast.bam']);
foreach my $input (@inputs) {
foreach my $format (@formats) {
# Only test -T option for now due to problems with reformatting
# the @SQ lines in the headers with embedded references.
foreach my $use_t (1) {
my @args = $use_t ? ('-T', $ref) : ();
if ($format->[0]) { push(@args, $format->[0]); }
push(@args, $input->[1]);
push(@args, '--no-PG');
my $compare = $format->[1] eq 'sam' ? 'compare' : 'compare_sam';
run_view_test($opts,
msg => "depad $format->[0] ($input->[0] input)",
cmd => 'depad',
args => \@args,
out => sprintf("%s.test%03d.%s",
$out, $test, $format->[1]),
ref_path => "$$opts{path}/dat/cram_md5",
redirect => 1,
$compare => $unpad_sam);
}
}
}
}
sub test_stats
{
my ($opts,%args) = @_;
my $efix = ($^O =~ /^(?:cygwin|msys|MSWin32)$/) ? 1 : 0;
test_cmd($opts,out=>'stat/1.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/1_map_cigar.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/1.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/1_map_cigar_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/2.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/2_equal_cigar_full_seq.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/2.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/2_equal_cigar_full_seq_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/3.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/3_map_cigar_equal_seq.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/3.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/3_map_cigar_equal_seq_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/4.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/4_X_cigar_full_seq.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/4.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/4_X_cigar_full_seq_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/5.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/5_insert_cigar.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/5.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/5_insert_cigar_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/6.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa -i 0 $$opts{path}/stat/5_insert_cigar.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/7.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/7_supp.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/7.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/7_supp_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/8.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/stat/test.fa $$opts{path}/stat/8_secondary.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/8.stats.large.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/8_secondary_large.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/9.stats.expected',cmd=>"$$opts{bin}/samtools stats -S RG -r $$opts{path}/stat/test.fa $$opts{path}/stat/1_map_cigar.sam | tail -n+4", exp_fix=>$efix,out_map=>{"stat/1_map_cigar.sam_s1_a_1.bamstat"=>"stat/1_map_cigar.sam_s1_a_1.expected.bamstat"},hskip=>3);
test_cmd($opts,out=>'stat/10.stats.expected',cmd=>"$$opts{bin}/samtools stats -S RG -r $$opts{path}/stat/test.fa $$opts{path}/stat/10_map_cigar.sam | tail -n+4", exp_fix=>$efix,out_map=>{"stat/10_map_cigar.sam_s1_a_1.bamstat"=>"stat/10_map_cigar.sam_s1_a_1.expected.bamstat", "stat/10_map_cigar.sam_s1_b_1.bamstat"=>"stat/10_map_cigar.sam_s1_b_1.expected.bamstat"},hskip=>3);
test_cmd($opts,out=>'stat/11.stats.expected',cmd=>"$$opts{bin}/samtools stats -t $$opts{path}/stat/11.stats.targets $$opts{path}/stat/11_target.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/11.stats.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/11_target.bam ref1:10-24 ref1:30-46 ref1:39-56 | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/11.stats.g4.expected',cmd=>"$$opts{bin}/samtools stats -g 4 -t $$opts{path}/stat/11.stats.targets $$opts{path}/stat/11_target.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/11.stats.g4.expected',cmd=>"$$opts{bin}/samtools stats -g 4 $$opts{path}/stat/11_target.bam ref1:10-24 ref1:30-46 ref1:39-56 | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/12.3reads.overlap.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/12_overlaps.bam -t $$opts{path}/stat/12_3reads.bed | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/12.3reads.nooverlap.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/12_overlaps.bam -p -t $$opts{path}/stat/12_3reads.bed | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/12.2reads.overlap.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/12_overlaps.bam -t $$opts{path}/stat/12_2reads.bed | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/12.2reads.nooverlap.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/12_overlaps.bam -p -t $$opts{path}/stat/12_2reads.bed | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/13.barcodes.bc.ok.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/13_barcodes_ok.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/13.barcodes.ox.ok.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/13_barcodes_ok_ox_bz.sam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/13.barcodes.fail.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/13_barcodes_fail_bc_length.sam | tail -n+4", expect_fail=>1, exp_fix=>$efix);
test_cmd($opts,out=>'stat/13.barcodes.fail.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/13_barcodes_fail_hyphen.sam | tail -n+4", expect_fail=>1, exp_fix=>$efix);
test_cmd($opts,out=>'stat/13.barcodes.fail.expected',cmd=>"$$opts{bin}/samtools stats $$opts{path}/stat/13_barcodes_fail_qt_length.sam | tail -n+4", expect_fail=>1, exp_fix=>$efix);
test_cmd($opts,out=>'stat/14.rg.s1.expected',cmd=>"$$opts{bin}/samtools stats -I s1 $$opts{path}/stat/11_target.bam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/14.rg.grp2.expected',cmd=>"$$opts{bin}/samtools stats -I grp2 $$opts{path}/stat/11_target.bam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/14.rg.grp3.expected',cmd=>"$$opts{bin}/samtools stats -I grp3 $$opts{path}/stat/11_target.bam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/14.rg.Sample.expected',cmd=>"$$opts{bin}/samtools stats -I Sample $$opts{path}/stat/11_target.bam | tail -n+4", exp_fix=>$efix);
test_cmd($opts,out=>'stat/15.stats.expected',cmd=>"$$opts{bin}/samtools stats -r $$opts{path}/mpileup/ce.fa $$opts{path}/stat/15.big_del.sam | tail -n+4", exp_fix=>$efix);
}
sub test_merge
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
# Note the use of -s 1 to fix the random seed in place
# Merge 1 - Standard 3 file SAM merge all presented on the command line
test_cmd($opts,out=>'merge/2.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -O sam - $$opts{path}/dat/test_input_1_a.sam $$opts{path}/dat/test_input_1_b.sam $$opts{path}/dat/test_input_1_c.sam");
# Merge 2 - Standard 3 file BAM merge all files presented on the command line
test_cmd($opts,out=>'merge/2.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -O sam - $$opts{path}/dat/test_input_1_a.bam $$opts{path}/dat/test_input_1_b.bam $$opts{path}/dat/test_input_1_c.bam");
# Merge 3 - Standard 3 file BAM merge 2 files in fofn 1 on command line
open(my $fofn, "$$opts{path}/merge/test_3.fofn");
my ($tmpfile_fh, $tmpfile_filename) = tempfile(UNLINK => 1);
while (<$fofn>) {
print $tmpfile_fh "$$opts{path}/$_";
}
close($tmpfile_fh);
test_cmd($opts,out=>'merge/3.merge.expected.sam', err=>'merge/3.merge.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -O sam -b $tmpfile_filename - $$opts{path}/dat/test_input_1_a.bam");
# Merge 4 - 1 file BAM merge with file presented on the command line
test_cmd($opts,out=>'merge/4.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -O sam - $$opts{path}/dat/test_input_1_b.bam");
# Merge 5 - 3 file SAM merge all presented on the command line override IDs to file names
test_cmd($opts,out=>'merge/5.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -r -s 1 -O sam - $$opts{path}/dat/test_input_1_a.sam $$opts{path}/dat/test_input_1_b.sam $$opts{path}/dat/test_input_1_c.sam");
# Merge 6 - merge all presented on the command line, combine PG and RG rather than dedup
test_cmd($opts,out=>'merge/6.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -cp -s 1 -O sam - $$opts{path}/dat/test_input_1_a.sam $$opts{path}/dat/test_input_1_b.sam");
# Merge 7 - ID and SN with regex in them
test_cmd($opts,out=>'merge/7.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -O sam - $$opts{path}/dat/test_input_1_a_regex.sam $$opts{path}/dat/test_input_1_b_regex.sam");
# Merge 8 - Standard 3 file SAM merge, output file specified via option
test_cmd($opts,out=>'merge/2.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -o - -s 1 -O sam $$opts{path}/dat/test_input_1_a.sam $$opts{path}/dat/test_input_1_b.sam $$opts{path}/dat/test_input_1_c.sam");
# Sort inputs by PG, then merge
system("$$opts{bin}/samtools sort -o $$opts{tmp}/merge.tag.1.bam -t PG -m 10M $$opts{path}/dat/test_input_1_b.sam") == 0 or die "failed to create sort BAM: $?";
system("$$opts{bin}/samtools sort -o $$opts{tmp}/merge.tag.2.bam -t PG -m 10M $$opts{path}/dat/test_input_1_d.sam") == 0 or die "failed to create sort BAM: $?";
test_cmd($opts,out=>'merge/tag.pg.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -p -c -t PG -O SAM - $$opts{tmp}/merge.tag.1.bam $$opts{tmp}/merge.tag.2.bam");
# Sort inputs by PG, then merge (name sorted)
system("$$opts{bin}/samtools sort -o $$opts{tmp}/merge.tag.3.bam -n -t PG -m 10M $$opts{path}/dat/test_input_1_c.sam") == 0 or die "failed to create sort BAM: $?";
system("$$opts{bin}/samtools sort -o $$opts{tmp}/merge.tag.4.bam -n -t PG -m 10M $$opts{path}/dat/test_input_1_d.sam") == 0 or die "failed to create sort BAM: $?";
test_cmd($opts,out=>'merge/tag.pg.n.merge.expected.sam', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools merge${threads} -s 1 -p -c -n -t PG -O SAM - $$opts{tmp}/merge.tag.3.bam $$opts{tmp}/merge.tag.4.bam");
# Check merge works when PG/RG/CO header lines absent (PR #1095)
# Note only "merges" one file, so expected output is input
test_cmd($opts, out=>'merge/test_no_pg_rg_co.sam', cmd => "$$opts{bin}/samtools merge${threads} --no-PG -O SAM - $$opts{path}/merge/test_no_pg_rg_co.sam");
system("$$opts{bin}/samtools view -ho $$opts{tmp}/merge.bed.1.bam --no-PG $$opts{path}/merge/merge.bed.1.sam") == 0 or die "failed to create merge.bed.1.bam: $?";
system("$$opts{bin}/samtools view -ho $$opts{tmp}/merge.bed.2.bam --no-PG $$opts{path}/merge/merge.bed.2.sam") == 0 or die "failed to create merge.bed.2.bam: $?";
system("$$opts{bin}/samtools index $$opts{tmp}/merge.bed.1.bam") == 0 or die "failed to index merge.bed.1.bam: $?";
system("$$opts{bin}/samtools index $$opts{tmp}/merge.bed.2.bam") == 0 or die "failed to index merge.bed.2.bam: $?";
test_cmd($opts, out=>'merge/merge.bed.expected.sam', cmd => "$$opts{bin}/samtools merge${threads} --no-PG -O SAM -L $$opts{path}/merge/merge.bed - $$opts{tmp}/merge.bed.1.bam $$opts{tmp}/merge.bed.2.bam");
# -r (RG from filename) option with no initial RG header
test_cmd($opts, out=>'merge/rg_from_r_mode.expected.sam', cmd => "$$opts{bin}/samtools merge${threads} --no-PG -r -O SAM - $$opts{path}/merge/test_no_pg_rg_co.sam");
# Check merge works for TemplateCoordinate
test_cmd($opts, out=>'merge/test_template_coordinate.expected.sam', cmd => "$$opts{bin}/samtools merge${threads} --no-PG -O SAM --template-coordinate - $$opts{path}/merge/test_template_coordinate.1.sam $$opts{path}/merge/test_template_coordinate.2.sam");
}
sub test_sort
{
my ($opts, %args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
# TODO Sort test cases
# Check obsolete invocation is detected
test_cmd($opts, out=>"dat/empty.expected", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} $$opts{path}/dat/test_input_1_a.bam $$opts{tmp}/sortout", want_fail=>1);
test_cmd($opts, out=>"dat/empty.expected", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -f $$opts{path}/dat/test_input_1_a.bam $$opts{tmp}/sortout.bam", want_fail=>1);
test_cmd($opts, out=>"dat/empty.expected", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -o $$opts{path}/dat/test_input_1_a.bam $$opts{tmp}/sorttmp", want_fail=>1);
# Pos sort
test_cmd($opts, out=>"sort/pos.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -m 10M $$opts{path}/dat/test_input_1_a.bam -O SAM -o -");
# Name sort
test_cmd($opts, out=>"sort/name.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -n -m 10M $$opts{path}/dat/test_input_1_a.bam -O SAM -o -");
test_cmd($opts, out=>"sort/name2.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -N -m 10M $$opts{path}/dat/test_input_1_b.bam -O SAM -o -");
test_cmd($opts, out=>"sort/name3.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -n -m 10M $$opts{path}/dat/sort_name_input_1.sam -O SAM -o -");
# Tag sort (RG)
test_cmd($opts, out=>"sort/tag.rg.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -t RG -m 10M $$opts{path}/dat/test_input_1_a.bam -O SAM -o -");
# Tag sort (RG); secondary by name
test_cmd($opts, out=>"sort/tag.rg.n.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -n -t RG -m 10M $$opts{path}/dat/test_input_1_a.bam -O SAM -o -");
# Tag sort (AS)
test_cmd($opts, out=>"sort/tag.as.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -t AS -m 10M $$opts{path}/dat/test_input_1_d.sam -O SAM -o -");
# Tag sort (FI)
test_cmd($opts, out=>"sort/tag.fi.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} -t FI -m 10M $$opts{path}/dat/test_input_1_d.sam -O SAM -o -");
# TemplateCoordinate sort
test_cmd($opts, out=>"sort/template-coordinate.sort.expected.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools sort${threads} --template-coordinate -m 10M $$opts{path}/sort/template-coordinate.sort.sam -O SAM -o -");
# Minimiser sort, basic
test_cmd($opts, out=>"sort/minimiser-basic.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools reset --dupflag $$opts{path}/dat/auto_indexed.tmp.bam | $$opts{bin}/samtools sort${threads} -m 10M -M -K10 -O SAM -o -");
# Minimiser sort, indexed reference
test_cmd($opts, out=>"sort/minimiser-indexed.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools reset --dupflag $$opts{path}/dat/auto_indexed.tmp.bam | $$opts{bin}/samtools sort${threads} -m 10M -M -K10 -I $$opts{path}/dat/mpileup.ref.fa -O SAM -o -");
# Minimiser sort, indexed reference plus homopolymer squash
test_cmd($opts, out=>"sort/minimiser-indexed-poly.sam", ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools reset --dupflag $$opts{path}/dat/auto_indexed.tmp.bam | $$opts{bin}/samtools sort${threads} -m 10M -MH -K10 -I $$opts{path}/dat/mpileup.ref.fa -O SAM -o -");
}
sub test_collate
{
my ($opts, %args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
# Output to stdout
test_cmd($opts, out=>"collate/collate.expected.sam", ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} --output-fmt=sam -O $$opts{path}/dat/test_input_1_d.sam");
# Output to file
test_cmd($opts, out=>"dat/empty.expected",
out_map=>{"collate/collate1.tmp.sam"
=>"collate/collate.expected.sam"}, ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} -o $$opts{path}/collate/collate1.tmp.sam $$opts{path}/dat/test_input_1_d.sam");
# Output to file, given tmp file prefix
test_cmd($opts, out=>"dat/empty.expected",
out_map=>{"collate/collate2.tmp.sam"
=>"collate/collate.expected.sam"}, ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} -o $$opts{path}/collate/collate2.tmp.sam $$opts{path}/dat/test_input_1_d.sam $$opts{path}/collate/collate2.tmp");
# Legacy usage with output file name based on prefix
test_cmd($opts, out=>"dat/empty.expected",
out_map=>{"collate/collate3.tmp.sam"
=>"collate/collate.expected.sam"}, ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} --output-fmt=sam $$opts{path}/dat/test_input_1_d.sam $$opts{path}/collate/collate3.tmp");
# fast collate, supplementary files not output
test_cmd($opts, out=>"dat/empty.expected",
out_map=>{"collate/1_fast_collate.sam" => "collate/1_fast_collate.sam.expected"}, ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} --output-fmt=sam -f $$opts{path}/collate/fast_collate.sam -o $$opts{path}/collate/1_fast_collate.sam");
# fast collate, supplementary files not output and force temp file
test_cmd($opts, out=>"dat/empty.expected",
out_map=>{"collate/2_fast_collate_with_tmp.sam" => "collate/2_fast_collate_with_tmp_used.sam.expected"}, ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools collate${threads} --output-fmt=sam -f -r 4 $$opts{path}/collate/fast_collate.sam -o $$opts{path}/collate/2_fast_collate_with_tmp.sam");
}
sub test_fixmate
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts,out=>'fixmate/1_coord_sort.sam.expected', err=>'fixmate/1_coord_sort.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -O sam $$opts{path}/fixmate/1_coord_sort.sam -", expect_fail=>1);
test_cmd($opts,out=>'fixmate/2_isize_overflow.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -z off -O sam $$opts{path}/fixmate/2_isize_overflow.sam -");
test_cmd($opts,out=>'fixmate/3_reverse_read_pp_lt.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -O sam $$opts{path}/fixmate/3_reverse_read_pp_lt.sam -");
test_cmd($opts,out=>'fixmate/4_reverse_read_pp_equal.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -O sam $$opts{path}/fixmate/4_reverse_read_pp_equal.sam -");
test_cmd($opts,out=>'fixmate/5_ct.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -cO sam $$opts{path}/fixmate/5_ct.sam -");
test_cmd($opts,out=>'fixmate/6_ct_replace.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -cO sam $$opts{path}/fixmate/6_ct_replace.sam -");
test_cmd($opts,out=>'fixmate/7_two_read_mapped.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -z off -O sam $$opts{path}/fixmate/7_two_read_mapped.sam -");
test_cmd($opts,out=>'fixmate/8_isize_overflow_64bit.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -z off -O sam $$opts{path}/fixmate/8_isize_overflow_64bit.sam -");
test_cmd($opts,out=>'fixmate/sanitize.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools fixmate${threads} -O sam $$opts{path}/fixmate/sanitize.sam -");
# fixmate -M base-modification tests
foreach (qw/ok+ ok- draft not_updated not_updated_noML not_updated_noMN
bad_MN MN_only ML_only ML_wrong_len noseq bounds/) {
test_cmd($opts,
out=>"fixmate/mod_$_.sam.expected",
ignore_pg_header => 1,
cmd=>"$$opts{bin}/samtools fixmate${threads} -M -z off -O sam $$opts{path}/fixmate/mod_$_.sam -");
}
}
sub test_reference
{
my ($opts,%args) = @_;
# Build a CRAM file with some missing data in the middle
cmd("$$opts{bin}/samtools view -e 'pos<1000||pos>1200' -O cram,embed_ref=1 -T test/dat/mpileup.ref.fa -o $$opts{path}/reference/mpileup.1.tmp.cram test/dat/mpileup.1.sam");
# MD:Z mode
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts,out=>'reference/mpileup.MD.fa.expected', cmd=>"$$opts{bin}/samtools reference${threads} test/reference/mpileup.1.tmp.cram");
# Embedded reference mode
test_cmd($opts,out=>'reference/mpileup.embed.fa.expected', cmd=>"$$opts{bin}/samtools reference${threads} -e test/reference/mpileup.1.tmp.cram");
# Region queries.
# Produce a region fasta file using faidx, and then supply this as
# the expected output to the reference command.
cmd("$$opts{bin}/samtools faidx test/reference/mpileup.MD.fa.expected");
cmd("$$opts{bin}/samtools faidx test/reference/mpileup.MD.fa.expected 17:1000-1500 > test/reference/mpileup.MD.fa.reg.tmp.expected");
cmd("$$opts{bin}/samtools faidx test/reference/mpileup.embed.fa.expected");
cmd("$$opts{bin}/samtools faidx test/reference/mpileup.embed.fa.expected 17:1000-1500 > test/reference/mpileup.embed.fa.reg.tmp.expected");
cmd("$$opts{bin}/samtools index test/reference/mpileup.1.tmp.cram");
test_cmd($opts,out=>'reference/mpileup.MD.fa.reg.tmp.expected', cmd=>"$$opts{bin}/samtools reference${threads} -r 17:1000-1500 test/reference/mpileup.1.tmp.cram");
test_cmd($opts,out=>'reference/mpileup.embed.fa.reg.tmp.expected', cmd=>"$$opts{bin}/samtools reference${threads} -r 17:1000-1500 -e test/reference/mpileup.1.tmp.cram");
}
sub test_calmd
{
my ($opts, %args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
my $test = "$$opts{bin}/samtools calmd${threads} -uAr $$opts{path}/dat/mpileup.1.sam $$opts{path}/dat/mpileup.ref.fa";
print "$test\n";
my $out = cmd($test);
if (substr($out, 0, 2) eq "\x1f\x8b") { passed($opts,msg=>$test); }
else { failed($opts,msg=>$test,reason=>"Expected BGZF-compressed output"); }
}
sub test_idxstat
{
my ($opts,%args) = @_;
test_cmd($opts,out=>'idxstats/test_input_1_a.bam.expected', err=>'idxstats/test_input_1_a.bam.expected.err', cmd=>"$$opts{bin}/samtools idxstats $$opts{path}/dat/test_input_1_a.bam", expect_fail=>0);
test_cmd($opts,out=>'idxstats/test_input_1_a.bam.expected', err=>'idxstats/test_input_1_a.bam.expected.err', cmd=>"$$opts{bin}/samtools idxstats $$opts{path}/dat/test_input_1_a.cram", expect_fail=>0);
test_cmd($opts,out=>'idxstats/test_input_1_a.bam.expected', err=>'idxstats/test_input_1_a.bam.expected.err', cmd=>"$$opts{bin}/samtools idxstats $$opts{path}/dat/test_input_1_a.sam", expect_fail=>0);
}
sub test_quickcheck
{
my ($opts,%args) = @_;
my @testfiles = (
'quickcheck/1.quickcheck.badeof.bam',
'quickcheck/2.quickcheck.badheader.bam',
'quickcheck/3.quickcheck.ok.bam',
'quickcheck/4.quickcheck.ok.bam',
'quickcheck/5.quickcheck.scramble30.truncated.cram',
'quickcheck/6.quickcheck.cram21.ok.cram',
'quickcheck/7.quickcheck.cram30.ok.cram',
'quickcheck/8.quickcheck.cram21.truncated.cram',
'quickcheck/9.quickcheck.cram30.truncated.cram',
'quickcheck/10.quickcheck.notargets.bam',
);
my $all_testfiles;
foreach my $fn (@testfiles) {
$all_testfiles .= " $$opts{path}/$fn";
test_cmd($opts, out => 'dat/empty.expected',
want_fail => ($fn !~ /[.]ok[.]/),
cmd => "$$opts{bin}/samtools quickcheck $$opts{path}/$fn");
}
test_cmd($opts, out => 'quickcheck/all.expected', want_fail => 1,
cmd => "$$opts{bin}/samtools quickcheck -v $all_testfiles | sed 's,.*/quickcheck/,,'");
test_cmd($opts, out => 'dat/empty.expected', want_fail => 0,
cmd => "$$opts{bin}/samtools quickcheck -uv $$opts{path}/quickcheck/10.quickcheck.notargets.bam | sed 's,.*/quickcheck/,,'");
}
sub test_reheader
{
my ($opts,%args) = @_;
local $ENV{REF_PATH} = "$$opts{path}/dat/cram_md5/%s";
my $fn = "$$opts{path}/dat/view.001";
# Create local BAM and CRAM inputs
system("$$opts{bin}/samtools view -b --no-PG $fn.sam > $fn.tmp.bam") == 0 or die "failed to create bam: $?";
system("$$opts{bin}/samtools view -C --no-PG --output-fmt-option version=2.1 $fn.sam > $fn.tmp.v21.cram") == 0 or die "failed to create cram: $?";
system("$$opts{bin}/samtools view -C --no-PG --output-fmt-option version=3.0 $fn.sam > $fn.tmp.v30.cram") == 0 or die "failed to create cram: $?";
# Fudge @PG lines. The version number will differ each commit.
# Also the pathname will differ for each install. We'll take it on faith
# that these bits work.
test_cmd($opts,
out=>'reheader/1_view1.sam.expected',
err=>'reheader/1_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader $$opts{path}/reheader/hdr.sam $fn.tmp.bam | $$opts{bin}/samtools view -h --no-PG | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
test_cmd($opts,
out=>'reheader/2_view1.sam.expected',
err=>'reheader/2_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader $$opts{path}/reheader/hdr.sam $fn.tmp.v21.cram | $$opts{bin}/samtools view -h --no-PG | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
test_cmd($opts,
out=>'reheader/2_view1.sam.expected',
err=>'reheader/2_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader $$opts{path}/reheader/hdr.sam $fn.tmp.v30.cram | $$opts{bin}/samtools view -h --no-PG | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
# In-place testing
test_cmd($opts,
out=>'reheader/3_view1.sam.expected',
err=>'reheader/3_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader --in-place $$opts{path}/reheader/hdr.sam $fn.tmp.v21.cram && $$opts{bin}/samtools view -h --no-PG $fn.tmp.v21.cram | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
test_cmd($opts,
out=>'reheader/3_view1.sam.expected',
err=>'reheader/3_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader --in-place $$opts{path}/reheader/hdr.sam $fn.tmp.v30.cram && $$opts{bin}/samtools view -h --no-PG $fn.tmp.v30.cram | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
test_cmd($opts,
out=>'reheader/4_view1.sam.expected',
err=>'reheader/1_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader -c \"sed \'s/2014 Genome/2019 Genome/g\'\" $fn.tmp.bam | $$opts{bin}/samtools view -h --no-PG | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
test_cmd($opts,
out=>'reheader/5_view1.sam.expected',
err=>'reheader/1_view1.sam.expected.err',
cmd=>"$$opts{bin}/samtools reheader -c \"sed \'s/2014 Genome/2019 Genome/g\'\" $fn.tmp.v30.cram | $$opts{bin}/samtools view -h --no-PG | perl -pe 's/\tVN:.*//'",
exp_fix=>1,
reorder_header => 1);
}
sub test_addrprg
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts,out=>'addrprg/1_fixup.sam.expected', err=>'addrprg/1_fixup.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -m overwrite_all $$opts{path}/addrprg/1_fixup.sam");
test_cmd($opts,out=>'addrprg/2_fixup_orphan.sam.expected', err=>'addrprg/2_fixup_orphan.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -m orphan_only $$opts{path}/addrprg/2_fixup_orphan.sam");
test_cmd($opts,out=>'addrprg/3_fixup.sam.expected', err=>'addrprg/3_fixup.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -R '1#9' $$opts{path}/addrprg/1_fixup.sam", want_fail=>1);
test_cmd($opts,out=>'addrprg/4_fixup_norg.sam.expected', err=>'addrprg/4_fixup_norg.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -r '\@RG\\tID:1#8\\tCN:SC' $$opts{path}/addrprg/4_fixup_norg.sam");
test_cmd($opts,out=>'addrprg/1_fixup.sam.expected', err=>'addrprg/1_fixup.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -m overwrite_all -R '1#8' $$opts{path}/addrprg/1_fixup.sam");
test_cmd($opts,out=>'addrprg/4_fixup_norg.sam.expected', err=>'addrprg/4_fixup_norg.sam.expected.err', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -r 'ID:1#8' -r 'CN:SC' $$opts{path}/addrprg/4_fixup_norg.sam");
test_cmd($opts,out=>'addrprg/5_editrg.sam.expected', ignore_pg_header => 1, cmd=>"$$opts{bin}/samtools addreplacerg${threads} -O sam -w -r '\@RG\\tID:1#8\\tCN:Sanger\\tDS:Testing the editing code.' $$opts{path}/addrprg/1_fixup.sam");
}
sub test_markdup
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts, out=>'markdup/1_name_sort.expected.sam', err=>'1_name_sort.expected.sam.err', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam --no-PG $$opts{path}/markdup/1_name_sort.sam -", expect_fail=>1);
test_cmd($opts, out=>'markdup/2_bad_order.expected.sam', err=>'2_bad_order.expected.sam.err', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam --no-PG $$opts{path}/markdup/2_bad_order.sam -", expect_fail=>1);
test_cmd($opts, out=>'markdup/3_missing_mc.expected.sam', err=>'3_missing_mc.expected.sam.err', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam --no-PG $$opts{path}/markdup/3_missing_mc.sam -", expect_fail=>1);
test_cmd($opts, out=>'markdup/4_missing_ms.expected.sam', err=>'4_missing_ms.expected.sam.err', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam --no-PG $$opts{path}/markdup/4_missing_ms.sam -", expect_fail=>1);
test_cmd($opts, out=>'markdup/5_markdup.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam --no-PG $$opts{path}/markdup/5_markdup.sam -");
test_cmd($opts, out=>'markdup/6_remove_dups.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -O sam -r --no-PG $$opts{path}/markdup/6_remove_dups.sam -");
test_cmd($opts, out=>'markdup/7_mark_supp_dup.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -O sam --no-PG $$opts{path}/markdup/7_mark_supp_dup.sam -");
test_cmd($opts, out=>'markdup/8_optical_dup.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t -O sam --no-PG $$opts{path}/markdup/8_optical_dup.sam -");
test_cmd($opts, out=>'markdup/9_optical_dup_qcfail.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 2500 --mode s -t --include-fails -O sam --no-PG $$opts{path}/markdup/9_optical_dup_qcfail.sam -");
test_cmd($opts, out=>'markdup/10_optical_chain.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 2500 --mode s -t -O sam --no-PG -S $$opts{path}/markdup/10_optical_chain.sam -");
test_cmd($opts, out=>'markdup/10_optical_chain.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 2500 --mode s -t -O sam --read-coords '([[:digit:]]+):([[:digit:]]+):([[:digit:]]+)\$' --no-PG -S $$opts{path}/markdup/10_optical_chain.sam -");
test_cmd($opts, out=>'markdup/11_optical_dup_regex.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t -O sam --read-coords '^([0-9]+):([0-9]+):([[:print:]]+)' --coords-order xyt --no-PG $$opts{path}/markdup/11_optical_dup_regex.sam -");
test_cmd($opts, out=>'markdup/11_optical_dup_regex.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t -O sam --read-coords '^([0-9]+):([0-9]+)' --coords-order xy --no-PG $$opts{path}/markdup/11_optical_dup_regex.sam -");
test_cmd($opts, out=>'markdup/12_optical_chain_regex.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 2500 --mode s -t -O sam --read-coords '([[:digit:]]+):([[:digit:]]+)\$' --coords-order xy --no-PG $$opts{path}/markdup/12_optical_chain_regex.sam -");
test_cmd($opts, out=>'markdup/13_optical_barcode_tag.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t --barcode-tag BX -O sam --no-PG $$opts{path}/markdup/13_optical_barcode_tag.sam -");
test_cmd($opts, out=>'markdup/14_optical_barcode_name.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t --barcode-name -O sam --no-PG $$opts{path}/markdup/14_optical_barcode_name.sam -");
test_cmd($opts, out=>'markdup/15_optical_barcode_rgx_name.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t --barcode-rgx '^([!-9;-?A-~]+):[0-9]+:' --read-coords '^[!-9;-?A-~]+:([0-9]+):([0-9]+)' --coords-order xy -O sam --no-PG $$opts{path}/markdup/15_optical_barcode_rgx_name.sam -");
test_cmd($opts, out=>'markdup/16_optical_barcode_rgx_name_test_2.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -S -d 100 --mode s -t --barcode-rgx '^([!-9;-?A-~]+):[0-9]+:' --read-coords '^[!-9;-?A-~]+:([0-9]{4})([0-9]{4})' --coords-order xy -O sam --no-PG $$opts{path}/markdup/16_optical_barcode_rgx_name_test_2.sam -");
test_cmd($opts, out=>'markdup/17_read_group.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} -d 100 --mode s -t --use-read-groups -O sam --no-PG $$opts{path}/markdup/17_read_group.sam -");
test_cmd($opts, out=>'markdup/18_primary_duplicate_count.expected.sam', cmd=>"$$opts{bin}/samtools markdup${threads} --mode t -t -O sam --no-PG --duplicate-count --barcode-tag BC -S $$opts{path}/markdup/18_primary_duplicate_count.sam -");
}
sub test_bedcov
{
my ($opts,%args) = @_;
my $out;
test_cmd($opts,out=>'bedcov/bedcov.expected',cmd=>"$$opts{bin}/samtools bedcov $$opts{path}/bedcov/bedcov.bed $$opts{path}/bedcov/bedcov.bam");
test_cmd($opts,out=>'bedcov/bedcov_j.expected',cmd=>"$$opts{bin}/samtools bedcov -j $$opts{path}/bedcov/bedcov.bed $$opts{path}/bedcov/bedcov.bam");
test_cmd($opts,out=>'bedcov/bedcov_gG.expected',cmd=>"$$opts{bin}/samtools bedcov -g512 -G2048 $$opts{path}/bedcov/bedcov_gG.bed $$opts{path}/bedcov/bedcov.bam");
test_cmd($opts,out=>'bedcov/bedcov_c.expected',cmd=>"$$opts{bin}/samtools bedcov -c $$opts{path}/bedcov/bedcov_gG.bed $$opts{path}/bedcov/bedcov.bam");
#with header
cmd("echo \"#chrom\tchromStart\tchromEnd\t$$opts{path}/bedcov/bedcov.bam_cov\" > $$opts{tmp}/bedcovH1.expected");
cmd ("cat $$opts{path}/bedcov/bedcov.expected >> $$opts{tmp}/bedcovH1.expected");
cmd("$$opts{bin}/samtools bedcov -H $$opts{path}/bedcov/bedcov.bed $$opts{path}/bedcov/bedcov.bam > $$opts{tmp}/out.H1");
$out = cmd("diff $$opts{tmp}/bedcovH1.expected $$opts{tmp}/out.H1");
if ( $out ne "" ) {
failed($opts,msg=>"coverage with header",reason=>"output does $out not match to expected\n");
} else {
passed($opts,msg=>"coverage with header success\n");
}
#with custom header
cmd("echo \"#chrom\tchromStart\tchromEnd\tT1\nchr1\t12209228\t12209246\t10\" > $$opts{tmp}/bedcovH2.bed");
cmd("echo \"#chrom\tchromStart\tchromEnd\tT1\t$$opts{path}/bedcov/bedcov.bam_cov\nchr1\t12209228\t12209246\t10\t24\" > $$opts{tmp}/bedcovH2.expected");
cmd("$$opts{bin}/samtools bedcov -H $$opts{tmp}/bedcovH2.bed $$opts{path}/bedcov/bedcov.bam > $$opts{tmp}/out.H2");
$out = cmd("diff $$opts{tmp}/bedcovH2.expected $$opts{tmp}/out.H2");
if ( $out ne "" ) {
failed($opts,msg=>"coverage with custom header",reason=>"output does not match to expected\n");
} else {
passed($opts,msg=>"coverage with custom header success\n");
}
#with empty source header
cmd("echo \"#chrom\tchromStart\tchromEnd\t\nchr1\t12209228\t12209246\t10\" > $$opts{tmp}/bedcovH3.bed");
cmd("echo \"#chrom\tchromStart\tchromEnd\t\t$$opts{path}/bedcov/bedcov.bam_cov\nchr1\t12209228\t12209246\t10\t24\" > $$opts{tmp}/bedcovH3.expected");
cmd("$$opts{bin}/samtools bedcov -H $$opts{tmp}/bedcovH3.bed $$opts{path}/bedcov/bedcov.bam > $$opts{tmp}/out.H3");
$out = cmd("diff $$opts{tmp}/bedcovH3.expected $$opts{tmp}/out.H3");
if ( $out ne "" ) {
failed($opts,msg=>"coverage with src empty header",reason=>"output does not match to expected\n");
} else {
passed($opts,msg=>"coverage with src empty header success\n");
}
#empty added header
#chrom\tchromStart\tchromEnd\tname\tscore\tstrand\tthickStart\t\
# thickEnd\titemRgb\tblockCount\tblockSizes\tblockStarts
cmd("echo \"chr1\t12209228\t12209246\t4\t5\t6\t7\t8\t9\t10\t11\t12\t13\t14\" > $$opts{tmp}/bedcovH4.bed");
cmd("echo \"#chrom\tchromStart\tchromEnd\tname\tscore\tstrand\tthickStart\tthickEnd\titemRgb\tblockCount\tblockSizes\tblockStarts\t.\t.\t$$opts{path}/bedcov/bedcov.bam_cov\" > $$opts{tmp}/bedcovH4.expected");
cmd("echo \"chr1\t12209228\t12209246\t4\t5\t6\t7\t8\t9\t10\t11\t12\t13\t14\t24\" >> $$opts{tmp}/bedcovH4.expected");
cmd("$$opts{bin}/samtools bedcov -H $$opts{tmp}/bedcovH4.bed $$opts{path}/bedcov/bedcov.bam > $$opts{tmp}/out.H4");
$out = cmd("diff $$opts{tmp}/bedcovH4.expected $$opts{tmp}/out.H4");
if ( $out ne "" ) {
failed($opts,msg=>"coverage with empty header",reason=>"output does not match to expected\n");
} else {
passed($opts,msg=>"coverage with empty header success\n");
}
}
sub test_split
{
my ($opts, %args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.0.sam' => 'split/split.expected.grp1.sam',
'split/split.tmp.1.sam' => 'split/split.expected.grp2.sam',
'split/split.tmp.unk.sam' => 'split/split.expected.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -u $$opts{path}/split/split.tmp.unk.sam -f $$opts{path}/split/split.tmp.\%#.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.00000.sam' => 'split/split.expected.grp1.sam',
'split/split.tmp.00001.sam' => 'split/split.expected.grp2.sam',
'split/split.tmp.unk.sam' => 'split/split.expected.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -u $$opts{path}/split/split.tmp.unk.sam -p 5 -f $$opts{path}/split/split.tmp.\%#.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.grp1.sam' => 'split/split.expected.grp1.sam',
'split/split.tmp.grp2.sam' => 'split/split.expected.grp2.sam',
'split/split.tmp.unk.sam' => 'split/split.expected.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -u $$opts{path}/split/split.tmp.unk.sam -f $$opts{path}/split/split.tmp.\%!.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.grp1.sam' => 'split/split.expected.grp1.sam',
'split/split.tmp.grp2.sam' => 'split/split.expected.grp2.sam',
'split/split.tmp.grp3.sam' => 'split/split.expected_d_RG.grp3.sam',
'split/split.tmp.unk.sam' => 'split/split.expected_d_RG.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -d RG -u $$opts{path}/split/split.tmp.unk.sam -f $$opts{path}/split/split.tmp.\%!.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.aardvark.sam' => 'split/split.expected_d_an.aardvark.sam',
'split/split.tmp.badger.sam' => 'split/split.expected_d_an.badger.sam',
'split/split.tmp.cat.sam' => 'split/split.expected_d_an.cat.sam',
'split/split.tmp.dog.sam' => 'split/split.expected_d_an.dog.sam',
'split/split.tmp.unk.sam' => 'split/split.expected_d_an.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -d an -u $$opts{path}/split/split.tmp.unk.sam -f $$opts{path}/split/split.tmp.\%!.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out=>"dat/empty.expected",
out_map => {
'split/split.tmp.badger.sam' => 'split/split.expected_d_an.badger.sam',
'split/split.tmp.cat.sam' => 'split/split.expected_d_an.cat.sam',
'split/split.tmp.dog.sam' => 'split/split.expected_d_an.dog.sam',
'split/split.tmp.unk.sam' => 'split/split.expected_d_an_M_3.unk.sam',
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -d an -M 3 -u $$opts{path}/split/split.tmp.unk.sam -f $$opts{path}/split/split.tmp.\%!.\%. $$opts{path}/split/split.sam");
test_cmd($opts,
out => "dat/empty.expected",
out_map => {
"split/split.tmp.d_nn.-2.sam" => "split/split.expected_d_nn.-2.sam",
"split/split.tmp.d_nn.-1.sam" => "split/split.expected_d_nn.-1.sam",
"split/split.tmp.d_nn.1.sam" => "split/split.expected_d_nn.1.sam",
"split/split.tmp.d_nn.2.sam" => "split/split.expected_d_nn.2.sam",
"split/split.tmp.d_nn.unk.sam" => "split/split.expected_d_nn.unk.sam",
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -f $$opts{path}/split/split.tmp.d_nn.\%!.\%. -d nn -u $$opts{path}/split/split.tmp.d_nn.unk.sam $$opts{path}/split/split_d_nn.sam");
test_cmd($opts,
out => "dat/empty.expected",
out_map => {
"split/split.tmp.d_nn.-0002.sam" => "split/split.expected_d_nn.-2.sam",
"split/split.tmp.d_nn.-0001.sam" => "split/split.expected_d_nn.-1.sam",
"split/split.tmp.d_nn.0001.sam" => "split/split.expected_d_nn.1.sam",
"split/split.tmp.d_nn.0002.sam" => "split/split.expected_d_nn.2.sam",
"split/split.tmp.d_nn.0unk.sam" => "split/split.expected_d_nn.unk.sam",
},
ignore_pg_header => 1,
reorder_header => 1,
cmd => "$$opts{bin}/samtools split $threads --output-fmt sam -f $$opts{path}/split/split.tmp.d_nn.\%!.\%. -p 4 -d nn -u $$opts{path}/split/split.tmp.d_nn.0unk.sam $$opts{path}/split/split_d_nn.sam");
}
sub test_ampliconclip
{
my ($opts,%args) = @_;
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts, out=>'ampliconclip/1_soft_clipped.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts,
out=>'ampliconclip/1_soft_clipped.expected.sam',
out_map => {
'ampliconclip/primer_counts.tsv' => 'ampliconclip/1_soft_clipped_primer_counts.expected.tsv',
},
cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --primer-counts $$opts{path}/ampliconclip/primer_counts.tsv --output-fmt=sam -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/1_hard_clipped.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --hard-clip -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts,
out=>'ampliconclip/1_soft_clipped_strand.expected.sam',
out_map => {
'ampliconclip/primer_counts.tsv' => 'ampliconclip/1_soft_clipped_strand_primer_counts.expected.tsv',
},
cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --strand --primer-counts $$opts{path}/ampliconclip/primer_counts.tsv -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/1_filter.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --strand --filter-len 185 -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/1_fail.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --strand --fail-len 185 -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/1_original_tag.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --original -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/1_delete_tag.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --output-fmt=sam -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/1_test_data.sam");
test_cmd($opts, out=>'ampliconclip/2_both_clipped.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --keep-tag --output-fmt=sam --strand --both-ends -b $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconclip/2_both_test_data.sam");
test_cmd($opts,
out=>'ampliconclip/3_multi_ref_clip.expected.sam',
out_map => {
'ampliconclip/primer_counts.tsv' => 'ampliconclip/3_multi_ref_data_primer_counts.expected.tsv',
},
cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --output-fmt=sam --keep-tag --primer-counts $$opts{path}/ampliconclip/primer_counts.tsv -b $$opts{path}/ampliconclip/multi_ref.bed $$opts{path}/ampliconclip/3_multi_ref_data.sam");
test_cmd($opts, out=>'ampliconclip/4_total_hc_data.expected.sam', cmd=>"$$opts{bin}/samtools ampliconclip${threads} --no-PG --output-fmt=sam --hard-clip -b $$opts{path}/ampliconclip/ac_test2.bed $$opts{path}/ampliconclip/4_total_hc_data.sam");
}
sub test_ampliconstats
{
my ($opts,%args) = @_;
my @inputs = ("$$opts{path}/ampliconclip/1_hard_clipped.expected.sam",
"$$opts{path}/ampliconclip/1_soft_clipped.expected.sam",
"$$opts{path}/ampliconclip/1_soft_clipped_strand.expected.sam",
"$$opts{path}/ampliconclip/2_both_clipped.expected.sam");
my $threads = exists($args{threads}) ? " -@ $args{threads}" : "";
test_cmd($opts, out=>'ampliconstats/stats.expected.txt', cmd=>"$$opts{bin}/samtools ampliconstats${threads} -S -t 50 -d 1,20,100 $$opts{path}/ampliconclip/ac_test.bed @inputs | grep -E -v 'Samtools version|Command line'");
test_cmd($opts, out=>'ampliconstats/stats_mixed.expected.txt', cmd=>"$$opts{bin}/samtools ampliconstats${threads} -c 0 $$opts{path}/ampliconclip/multi_ref.bed $$opts{path}/ampliconstats/mixed_clipped.sam | grep -E -v 'Samtools version|Command line'");
test_cmd($opts, out=>'ampliconstats/stats_partial.expected.txt', cmd=>"$$opts{bin}/samtools ampliconstats${threads} -c 0 $$opts{path}/ampliconclip/ac_test.bed $$opts{path}/ampliconstats/mixed_clipped.sam | grep -E -v 'Samtools version|Command line'");
}
sub test_reset
{
my ($opts, %args) = @_;
#some tests uses samtools view to skip the header and check data part
#basic op 1, from pipe, to std out
test_cmd($opts, out=>"reset/basic.1.mp.1.expected", err=>"reset/empty.expected", cmd=>"cat $$opts{bin}/test/dat/mpileup.1.sam | $$opts{bin}/samtools reset --dupflag | $$opts{bin}/samtools view");
#basic op 2, from redirection
test_cmd($opts, out=>"reset/basic.1.mp.1.expected", err=>"reset/empty.expected", cmd=>"$$opts{bin}/samtools reset --dupflag < $$opts{bin}/test/dat/mpileup.1.sam | $$opts{bin}/samtools view");
#basic op 3, explicit input
test_cmd($opts, out=>"reset/basic.1.mp.1.expected", err=>"reset/empty.expected", cmd=>"$$opts{bin}/samtools reset --dupflag $$opts{bin}/test/dat/mpileup.1.sam | $$opts{bin}/samtools view");
#basic op 4, output to given file
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, out_map=>{"reset/output.tmp.sam" => 'reset/basic.output.mp.1.expected'}, ignore_pg_header=>1, cmd=>"cat $$opts{bin}/test/dat/mpileup.1.sam | $$opts{bin}/samtools reset --dupflag -o $$opts{bin}/test/reset/output.tmp.sam");
#basic op 5, bam input
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, out_map=>{"reset/output.tmp.sam" => 'reset/basic.bam.input.expected'}, ignore_pg_header=>1, cmd=>"$$opts{bin}/samtools reset --dupflag -o $$opts{bin}/test/reset/output.tmp.sam $$opts{bin}/test/dat/test_input_1_a.bam");
#basic op 6, cram input
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, out_map=>{"reset/output.tmp.sam" => 'reset/basic.cram.input.expected'}, ignore_pg_header=>1, cmd=>"$$opts{bin}/samtools reset --dupflag -o $$opts{bin}/test/reset/output.tmp.sam $$opts{bin}/test/dat/test_input_1_a.cram");
#reject-PG 1, reject all
test_cmd($opts, out=>"reset/reject.1.expected", err=>"reset/empty.expected", cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam -o $$opts{bin}/test/reset/output.tmp.sam; grep $$opts{bin}/test/reset/output.tmp.sam -e\"\@PG\tID\:samtools\tPN\:samtools\" | wc -l | sed 's/ //g'");
#reject-PG 2 keep bwa and remove samtools onwards
test_cmd($opts, out=>"reset/reject.2.expected", err=>"reset/empty.expected", cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG sam_to_fixed_bam $$opts{bin}/test/dat/mpileup.1.sam -o $$opts{bin}/test/reset/output.tmp.sam; grep $$opts{bin}/test/reset/output.tmp.sam -e\"\@PG\tID\:\" | wc -l | sed 's/ //g'");
#no-PG, grep should fail as no PG entry found
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", want_fail=>1, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index --no-PG $$opts{bin}/test/dat/mpileup.1.sam -o $$opts{bin}/test/reset/output.tmp.sam; grep $$opts{bin}/test/reset/output.tmp.sam -e\"\@PG\"");
#no-RG, no @RG
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.nRG.1.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG -o $$opts{bin}/test/reset/output");
#no-RG, no RG:Z even with keep-tag
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.nRG.2.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG --keep-tag RG -o $$opts{bin}/test/reset/output");
#keep tag, X0 and MD
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.keep.1.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG --keep-tag X0,MD -o $$opts{bin}/test/reset/output");
#override remove
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.keep.1.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG --remove-tag X0,X1,MD --keep-tag X0,MD -o $$opts{bin}/test/reset/output");
#remove tag
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.keep.2.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG --remove-tag X0,X1,MD -o $$opts{bin}/test/reset/output");
#remove through x
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.keep.2.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG -x X0,X1,MD -o $$opts{bin}/test/reset/output");
#keep through remove/^ + X1
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.keep.3.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag --reject-PG bwa_index $$opts{bin}/test/dat/mpileup.1.sam --no-RG --remove-tag ^X0,MD --keep-tag X1 -o $$opts{bin}/test/reset/output");
#flag update and reverse flip
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.flg.1.expected"}, cmd=>"$$opts{bin}/samtools reset --dupflag $$opts{bin}/test/reset/seq.sam -o $$opts{bin}/test/reset/output");
#flag update default
test_cmd($opts, out=>"reset/empty.expected", err=>"reset/empty.expected", hskip=>1, ignore_pg_header=>1, out_map=>{"reset/output" => "reset/output.flg.2.expected"}, cmd=>"$$opts{bin}/samtools reset $$opts{bin}/test/reset/seq.sam -o $$opts{bin}/test/reset/output");
}
sub test_checksum
{
my ($opts, %args) = @_;
my $chk = exists($args{threads}) ?"checksum -@ $args{threads}" :"checksum";
# Basic mode, two files with one read-group removed and subtle QC diffs
# 1.1 and 2.1 checksums should have the same "all" and ERR013140 and
# ERR156632 groups should match, with "-" matching delted ERR016352.
# tr -d '\011' is to fix windows nl-cr endings which break the comparison.
test_cmd($opts, out=>"checksum/chk1.1.expected", cmd=>"$$opts{bin}/samtools $chk $$opts{path}/checksum/chk1.bam | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
test_cmd($opts, out=>"checksum/chk2.1.expected", cmd=>"$$opts{bin}/samtools $chk $$opts{path}/checksum/chk2.cram | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
# Whole file comparison.
# These checksums differ to above as the order is now included
test_cmd($opts, out=>"checksum/chk2.2.expected", cmd=>"$$opts{bin}/samtools $chk -a $$opts{path}/checksum/chk2.cram | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
# QC fail and verbose modes
test_cmd($opts, out=>"checksum/chk2.3.expected", cmd=>"$$opts{bin}/samtools $chk -qv $$opts{path}/checksum/chk2.cram | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
test_cmd($opts, out=>"checksum/chk2.4.expected", cmd=>"$$opts{bin}/samtools $chk -qv -a $$opts{path}/checksum/chk2.cram | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
# Splits and merging.
# Basic mode
cmd("$$opts{bin}/samtools split -f '$$opts{path}/checksum/chk1-%!.tmp' $$opts{path}/checksum/chk1.bam");
foreach my $rg (qw/ERR013140 ERR016352 ERR156632/) {
cmd("$$opts{bin}/samtools checksum $$opts{path}/checksum/chk1-$rg.tmp -o $$opts{path}/checksum/chk1-$rg.tmp.chk");
}
test_cmd($opts, out=>"checksum/chk1.1.expected", cmd=>"$$opts{bin}/samtools $chk -m $$opts{path}/checksum/chk1-ERR*.tmp.chk | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
# Full alignment mode
cmd("$$opts{bin}/samtools split -u $$opts{path}/checksum/chk2-noRG.tmp -f '$$opts{path}/checksum/chk2-%!.tmp' $$opts{path}/checksum/chk2.cram");
foreach my $rg (qw/ERR013140 ERR156632 noRG/) {
cmd("$$opts{bin}/samtools checksum -a $$opts{path}/checksum/chk2-$rg.tmp -o $$opts{path}/checksum/chk2-$rg.tmp.chk");
}
test_cmd($opts, out=>"checksum/chk2.2.expected", cmd=>"$$opts{bin}/samtools $chk -m $$opts{path}/checksum/chk2-*.tmp.chk | sed 's/\\(# Checksum[^:]*:\\).*/\\1/'");
}
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