File: inspect.pl

package info (click to toggle)
bowtie 1.3.1-3
  • links: PTS, VCS
  • area: main
  • in suites: forky, sid, trixie
  • size: 15,816 kB
  • sloc: cpp: 37,094; perl: 5,806; ansic: 1,474; sh: 1,197; python: 462; makefile: 419
file content (377 lines) | stat: -rwxr-xr-x 10,227 bytes parent folder | download | duplicates (3)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
##
# inspect.pl
#
# Basic tests to ensure bowtie-inspect is behaving as expected.
#
# OLD_BOWTIE=~/software/bowtie-0.12.5 && \
# perl scripts/test/inspect.pl \
#    -bowtie-build2=$OLD_BOWTIE/bowtie-build \
#    -bowtie-inspect2=$OLD_BOWTIE/bowtie-inspect
#

use strict;
use warnings;
use Getopt::Long;

my $bowtie = "./bowtie";
my $bowtie2 = "./bowtie";
my $bowtie_build = "./bowtie-build";
my $bowtie_build2 = "./bowtie-build";
my $bowtie_inspect = "./bowtie-inspect";
my $bowtie_inspect2 = "./bowtie-inspect";
my $ref = "";
my $seed = 0;
my $debug = 0;
my $debug_old = 0;
my $nodie = 0;

GetOptions (
	"ref:s"                => \$ref,
	"bowtie:s"             => \$bowtie,
	"bowtie2:s"            => \$bowtie2,
	"bowtie-build:s"       => \$bowtie_build,
	"bowtie-build2:s"      => \$bowtie_build2,
	"bowtie-inspect:s"     => \$bowtie_inspect,
	"bowtie-inspect2:s"    => \$bowtie_inspect2,
	"seed:i"               => \$seed,
	"nodie"                => \$nodie,
	"debug"                => \$debug,
	"debug2"               => \$debug_old);

my $bowtie_d = "${bowtie} --debug";
my $bowtie2_d = "${bowtie2} --debug";
my $bowtie_build_d = "${bowtie_build} --debug";
my $bowtie_build2_d = "${bowtie_build2} --debug";
my $bowtie_inspect_d = "${bowtie_inspect} --debug";
my $bowtie_inspect2_d = "${bowtie_inspect2} --debug";

my @cases = (
	">\nN\n>0\nATCTAG\n>\nN\n",
	">-\nNA\n",
	">-\nNNA\n",
	">-\nNAA\n",
	">\nN\n>0\nATCTAG\n>\nN\n",
	">a\nN\n>b\nATCTAG\n>c\nN\n>d\nATCTAG\n",
	">a\nN\n>b\nATCTAG\n>c\nN\n",
	">\nN\n>0\nATCTAG\n>\nN\n",
	">a\nN\n>a\nNNNATCTAGNN\n>b\nNNN\n>c\nNNNNNN\n>d\nNNNNNA\n",
	">a\nNA\n>zzz\nNNNATCTAGNN\n>b\nNNN\n>c\nNNNNNN\n>d\nNA\n",
	">a\nAN\n>a\nNNNATCTAGNNA\n>b\nNNN\n>a\nNNNNNN\n>d\nAN\n"
);

sub mydie($) {
	my $msg = shift;
	print STDERR "$msg\n";
	if($nodie) {
		print "Press enter to continue...";
		my $tmp = <STDIN>;
	} else {
		die;
	}
}

##
# Given a fasta file string, strip away all sequences that consist
# entirely of gaps.
#
sub stripAllGaps($$) {
	my ($fa, $col) = @_;
	my @l = split(/[\r\n]+/, $fa);
	my $ret = "";
	while(1) {
		my $name = shift @l;
		last unless defined($name);
		substr($name, 0, 1) eq ">" || mydie("Unexpected name line:\n$name\nin fasta:\n$_[0]");
		my $seq = shift @l;
		if($col) {
			if($seq =~ /[ACGT][ACGT]/i) {
				$ret .= "$name\n$seq\n";
			}
		} else {
			if($seq =~ /[ACGT]/i) {
				$ret .= "$name\n$seq\n";
			}
		}
	}
	return $ret;
}

##
# Given a string in nucleotide space, convert to colorspace.
#
sub colorize($$) {
	my ($s, $nucs) = @_;
	defined($s) || die;
	my %cmap = (
		"AA" => "0", "CC" => "0", "GG" => "0", "TT" => "0",
		"AC" => "1", "CA" => "1", "GT" => "1", "TG" => "1",
		"AG" => "2", "GA" => "2", "CT" => "2", "TC" => "2",
		"AT" => "3", "TA" => "3", "CG" => "3", "GC" => "3",
		"NA" => ".", "NC" => ".", "NG" => ".", "NT" => ".",
		"AN" => ".", "CN" => ".", "GN" => ".", "TN" => ".",
		"NN" => "."
	);
	my %nmap = ("0" => "A", "1" => "C", "2" => "G", "3" => "T", "." => "N");
	my $ret = "";
	for(my $i = 0; $i < length($s)-1; $i++) {
		my $di = uc substr($s, $i, 2);
		$di =~ tr/-NnMmRrWwSsYyKkVvHhDdBbXx/N/;
		defined($cmap{$di}) || mydie("Bad dinuc: $di\n");
		$ret .= ($nucs ? $nmap{$cmap{$di}} : $cmap{$di});
	}
	return $ret;
}

##
# Return version of argument with whitespace trimmed off either end.
#
sub trim($) {
	my $ret = $_[0];
	$ret =~ s/^\s+//; $ret =~ s/\s+$//;
	return $ret;
}

##
# Return version of argument with given character trimmed off either end.
#
sub trimc($$) {
	my $ret = $_[0];
	my $c = $_[1];
	$ret =~ s/^$c+//; $ret =~ s/$c+$//;
	return $ret;
}

##
# Given a fasta string of references, parse out the sequences and
# return them joined with commas.
#
sub refsToReads($$) {
	my ($fa, $col) = @_;
	my @l = split(/[\r\n]+/, $fa);
	my $fn = ".inspect.pl.refsToReads.fa";
	open(OUT, ">$fn") || die;
	while(1) {
		my $name = shift @l;
		last unless defined($name);
		my $seq = shift @l;
		$seq = trimc($seq, 'N');
		$seq = colorize($seq, 1) if $col;
		$seq =~ s/N.*$//i;
		$seq !~ /N/i || die;
		if(length($seq) > 0) {
			print OUT "$name\n$seq\n";
		}
	}
	close(OUT);
	return $fn;
}

##
# Given a fasta string or references and a hash ref, fill the hash ref
# with with reference sequences keyed by name.
#
sub refmap($$$) {
	my ($fa, $map, $col) = @_;
	my @l = split(/[\r\n]+/, $fa);
	my $namelessref = 0;
	while(1) {
		my $name = shift @l;
		last unless defined($name);
		$name = substr($name, 1);
		next if scalar(@l) == 0 || $l[0] =~ /^>/;
		my $seq = shift @l;
		next if !defined($seq) || length($seq) == 0;
		if($name eq "") {
			$name = $namelessref;
			$namelessref++;
		}
		my $trimseq = trimc($seq, 'N');
		$trimseq =~ s/N.*$//i;
		$trimseq !~ /N/i || die;
		if($col) {
			$trimseq = "" unless $trimseq =~ /[acgt][acgt]/i;
		}
		$map->{seq}->{$name}->{$seq}++ if $seq ne "";
		$map->{trimseq}->{$name}->{$trimseq}++ if $trimseq ne "";
		$map->{trimseq}->{$name}{str} = $trimseq if $trimseq ne "";
	}
}

##
# Given output from running bowtie (first arg) and the fasta string of
# references (second), make sure that every reference is represented
# at least once in the bowtie output.  (at least b/c some references
# might be substrings of others.)
#
sub reconcileAlsWithRefs($$$) {
	my ($als, $fa, $col) = @_;
	my %rm = ();
	refmap($fa, \%rm, $col);
	return if $col;
	# Make a map from ref strs to the alignments that mapped
	my @l = split(/[\r\n]+/, $als);
	my %hits = ();
	for my $al (@l) {
		my @ls = split(/\t/, $al);
		my ($rd, $fw, $ref, $off, $seq) = ($ls[0], $ls[1], $ls[2], $ls[3], $ls[4]);
		$hits{$ref} = $seq if ($rd eq $ref && $fw eq '+');
	}
	for my $i (keys %{$rm{trimseq}}) {
		defined($hits{$i}) || mydie("No hit for reference $i:\n$als");
		defined($rm{trimseq}{$i}{$hits{$i}}) ||
			mydie("Hit sequence:\n\"$hits{$i}\"\n doesn't match ref ".
			      "sequence:\n\"".$rm{trimseq}{$i}{str}."\"\n$als");
	}
}

##
# Check that two strings are the same and, if they're not, print the
# provided error message along with the output of a 'diff' between
# them.
#
sub match($$$) {
	if($_[0] ne $_[1]) {
		open(D1, ">.inspect.pl.$seed.d1") || die;
		open(D2, ">.inspect.pl.$seed.d2") || die;
		print D1 "$_[0]\n";
		print D2 "$_[1]\n";
		close(D1);
		close(D2);
		system("diff -uw .inspect.pl.$seed.d1 .inspect.pl.$seed.d2");
		mydie("$_[2]");
	}
}

##
# Given a fasta file string, strip away all sequences that consist
# entirely of gaps.
#
sub colorizeFasta($) {
	my @l = split(/[\r\n]+/, $_[0]);
	my $ret = "";
	while(1) {
		my $name = shift @l;
		last unless defined($name);
		substr($name, 0, 1) eq ">" || die;
		my $seq = shift @l;
		my $cseq = colorize($seq, 1);
		$ret .= "$name\n$cseq\n" if $cseq ne "";
	}
	return $ret;
}

if($ref ne "") {
	open(REF, $ref) || mydie("Could not open -ref $ref");
	@cases = ();
	push @cases, "";
	while(<REF>) {
		chomp;
		unless(/^>/) { s/[^ACGT]/N/gi; }
		$cases[0] .= "$_\n";
	}
	close(REF);
}

my $fn = ".inspect.pl.tmp.$seed.fa";
for my $ca (@cases) {
	for my $col (0,) {
		my ($c, $e) = split(/:/, $ca);
		if(stripAllGaps($c, $col) eq "") {
			print "Skipping test case because it had no unambiguous stretches\n";
			next;
		}
		open(TMP, ">$fn") || die;
		print TMP $c;
		close(TMP);
		my $bb = $debug ? $bowtie_build_d : $bowtie_build;
		$bb .= " -C" if $col;
		my $bi = $debug ? $bowtie_inspect_d : $bowtie_inspect;
		$bi .= " -a -1";
		if($bowtie_build2 ne "") {
			my $cmdEnd = "$fn $fn >/dev/null && $bowtie_inspect -s --extra $fn | grep -v '^Sequence' | grep -v 'refnames.size' | grep -v 'Reverse'";
			my $bbo = $debug_old ? $bowtie_build2_d : $bowtie_build2;
			$bbo .= " -C" if $col;
			my $cmd = "$bbo $cmdEnd";
			print "$cmd\n";
			my $bb2out = `$cmd`;
			print "Old bowtie-build:\n$bb2out\n";
			$cmd = "$bb $cmdEnd";
			print "$cmd\n";
			my $bbout = `$cmd`;
			print "New bowtie-build:\n$bbout\n";
			match($bbout, $bb2out, "RefRecords from new and old bowtie-build (above) didn't match");
		}
		my $cmd = "$bb $fn $fn >/dev/null";
		print "$cmd\n";
		system($cmd) == 0 || mydie("Exitlevel $? from command '$cmd'");
		$cmd = "$bi $fn";
		print "$cmd\n";
		my $io = trim(`$cmd`);
		$? == 0 || mydie("Exitlevel $? from command '$cmd'");
		my $msg = "Output from bowtie-inspect:\n$io\ndidn't match input:\n";
		if(defined($e)) {
			my $e2 = trim(stripAllGaps($e, $col));
			match($io, $e2, "$msg$e2");
		} else {
			my $c2 = trim(stripAllGaps($c, $col));
			match($io, $c2, "$msg$c2");
		}
		print "$io\n";
		$cmd = "$bi -e $fn";
		print "$cmd\n";
		$io = trim(`$cmd`);
		$? == 0 || mydie("Exitlevel $? from command '$cmd'");
		$msg = "Output from bowtie-inspect -e:\n$io\ndidn't match input:\n";
		if(defined($e)) {
			# Colorspace quandry: strip gaps then colorize, or vice versa?
			my $e2 = ($col ? colorizeFasta($e) : $e);
			$e2 = trim(stripAllGaps($e2, 0));
			match($io, $e2, "$msg$e2");
		} else {
			my $c2 = ($col ? colorizeFasta($c) : $c);
			$c2 = trim(stripAllGaps($c2, 0));
			match($io, $c2, "$msg$c2");
		}
		print "$io\n";
		if($bowtie_inspect2 ne "") {
			my $bio = $debug_old ? $bowtie_inspect2_d : $bowtie_inspect2;
			$bio .= " -a -1";
			$cmd = "$bio $fn";
			print "$cmd\n";
			$io = trim(`$cmd`);
			$? == 0 || mydie("Exitlevel $? from command '$cmd'");
			$msg = "Output from bowtie-inspect:\n$io\ndidn't match input:\n";
			if(defined($e)) {
				my $e2 = trim(stripAllGaps($e, $col));
				match($io, $e2, "$msg$e2");
			} else {
				my $c2 = trim(stripAllGaps($c, $col));
				match($io, $c2, "$msg$c2");
			}
			print "$io\n";
		}
		
		#
		# Now query the index using all of the reference strings as
		# queries and using '-a -v 0'.  This will test whether bowtie
		# (and potentially also an older version of bowtie) agrees with
		# bowtie-inspect and bowtie-build about what's in the index.
		#
		my $ba = $debug ? $bowtie_d : $bowtie;
		$ba .= " -C --col-keepends" if $col;
		$cmd = "$ba -f -a -v 0 $fn ".refsToReads($c, $col);
		print "$cmd\n";
		my $bao = trim(`$cmd`);
		reconcileAlsWithRefs($bao, $c, $col);
		if($bowtie2 ne "") {
			my $ba2 = $debug ? $bowtie2_d : $bowtie2;
			$ba2 .= " -C --col-keepends" if $col;
			$cmd = "$ba2 -f -a -v 0 $fn ".refsToReads($c, $col);
			print "$cmd\n";
			my $bao2 = trim(`$cmd`);
			reconcileAlsWithRefs($bao2, $c, $col);
		}
		
		print "PASSED\n";
	}
}