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 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804 805 806 807 808 809 810 811 812 813 814 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843 844 845 846 847 848 849 850 851 852 853 854 855 856 857 858 859 860 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880 881 882 883 884 885 886 887 888 889 890 891 892 893 894 895 896 897 898 899 900 901 902 903 904 905 906 907 908 909 910 911 912
|
#!/usr/bin/env perl
#
# Copyright 2010-2016, Julian Catchen <jcatchen@illinois.edu>
#
# This file is part of Stacks.
#
# Stacks is free software: you can redistribute it and/or modify
# it under the terms of the GNU General Public License as published by
# the Free Software Foundation, either version 3 of the License, or
# (at your option) any later version.
#
# Stacks is distributed in the hope that it will be useful,
# but WITHOUT ANY WARRANTY; without even the implied warranty of
# MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
# GNU General Public License for more details.
#
# You should have received a copy of the GNU General Public License
# along with Stacks. If not, see <http://www.gnu.org/licenses/>.
#
#
# Process the data for a genetic map: build stacks in parents and progeny,
# create a catalog from the parents, and match progeny against the catatlog.
# Call genotypes, and load all data into an MySQL database along the way.
#
# For the database interactions to work, the 'mysql' program is expected to be
# on the path and sufficient permissions set to access the specified database.
#
use strict;
use POSIX;
use File::Temp qw/ mktemp /;
use File::Spec;
use constant stacks_version => "_VERSION_";
use constant true => 1;
use constant false => 0;
my $dry_run = false;
my $sql = true;
my $create_db = false;
my $overw_db = false;
my $mysql_config = "_PKGDATADIR_" . "sql/mysql.cnf";
my $mysql_tables = "_PKGDATADIR_" . "sql/stacks.sql";
my $exe_path = "_BINDIR_";
my $out_path = "";
my $popmap_path = "";
my $sample_path = "";
my $db = "";
my $data_type = "map";
my $min_cov = 0;
my $min_rcov = 0;
my $batch_id = -1;
my $sample_id = 1;
my $desc = ""; # Database description of this dataset
my $date = ""; # Date relevent to this data, formatted for SQL: 2009-05-31
my $gzip = false;
my @parents;
my @progeny;
my @samples;
my (@_pstacks, @_cstacks, @_sstacks, @_genotypes, @_populations);
my $cmd_str = $0 . " " . join(" ", @ARGV);
parse_command_line();
my $cnf = (-e $ENV{"HOME"} . "/.my.cnf") ? $ENV{"HOME"} . "/.my.cnf" : $mysql_config;
#
# Check for the existence of the necessary pipeline programs
#
die ("Unable to find '" . $exe_path . "pstacks'.\n") if (!-e $exe_path . "pstacks" || !-x $exe_path . "pstacks");
die ("Unable to find '" . $exe_path . "cstacks'.\n") if (!-e $exe_path . "cstacks" || !-x $exe_path . "cstacks");
die ("Unable to find '" . $exe_path . "sstacks'.\n") if (!-e $exe_path . "sstacks" || !-x $exe_path . "sstacks");
die ("Unable to find '" . $exe_path . "genotypes'.\n") if (!-e $exe_path . "genotypes" || !-x $exe_path . "genotypes");
die ("Unable to find '" . $exe_path . "populations'.\n") if (!-e $exe_path . "populations" || !-x $exe_path . "populations");
die ("Unable to find '" . $exe_path . "index_radtags.pl'.\n") if (!-e $exe_path . "index_radtags.pl" || !-x $exe_path . "index_radtags.pl");
my ($log, $log_fh, $sample);
my (@sample_list, %pop_ids, %pops, %grp_ids, %grps, %sample_ids);
parse_population_map(\@sample_list, \%pop_ids, \%pops, \%grp_ids, \%grps);
initialize_samples(\@parents, \@progeny, \@samples, \@sample_list, \%pop_ids, \%grp_ids);
#
# Open the log file
#
$log = "$out_path/ref_map.log";
open($log_fh, ">$log") or die("Unable to open log file '$log'; $!\n");
print $log_fh
"ref_map.pl version ", stacks_version, " started at ", strftime("%Y-%m-%d %H:%M:%S", (localtime(time))), "\n",
$cmd_str, "\n\n";
initialize_database($log_fh, \@parents, \@progeny, \@samples, \%sample_ids) if ($sql == true);
execute_stacks($log_fh, $sample_id, \@parents, \@progeny, \@samples, \%sample_ids);
load_sql_data($log_fh, \%pops, \@parents, \@progeny, \@samples) if ($sql == true);
print $log_fh "\nref_map.pl completed at ", strftime("%Y-%m-%d %H:%M:%S", (localtime(time))), "\n";
close($log_fh);
sub execute_stacks {
my ($log_fh, $sample_id, $parents, $progeny, $samples, $sample_ids) = @_;
my (@results, @depths_of_cov);
my ($pop_cnt, $sample, $num_files, $i, $cmd, $pipe_fh, $path, $cat_file);
my $minc = $min_cov > 0 ? "-m $min_cov" : "";
my $minrc = $min_rcov > 0 ? "-m $min_rcov" : $minc;
$i = 1;
$num_files = scalar(@{$parents}) + scalar(@{$progeny}) + scalar(@{$samples});
#
# Assemble RAD loci in each individual.
#
foreach $sample (@parents, @progeny, @samples) {
printf("Identifying unique stacks; file % 3s of % 3s [%s]\n", $i, $num_files, $sample->{'file'});
printf($log_fh "Identifying unique stacks; file % 3s of % 3s [%s]\n", $i, $num_files, $sample->{'file'});
if (scalar(keys %{$sample_ids}) > 0) {
$sample_id = $sample_ids->{$sample->{'file'}};
}
$path = $sample->{'path'} . $sample->{'file'} . "." . $sample->{'suffix'};
if ($sample->{'type'} eq "sample") {
$cmd = $exe_path . "pstacks -t $sample->{'fmt'} -f $path -o $out_path -i $sample_id $minc " . join(" ", @_pstacks) . " 2>&1";
} elsif ($sample->{'type'} eq "parent") {
$cmd = $exe_path . "pstacks -t $sample->{'fmt'} -f $path -o $out_path -i $sample_id $minc " . join(" ", @_pstacks) . " 2>&1";
} elsif ($sample->{'type'} eq "progeny") {
$cmd = $exe_path . "pstacks -t $sample->{'fmt'} -f $path -o $out_path -i $sample_id $minrc " . join(" ", @_pstacks) . " 2>&1";
}
print STDERR " $cmd\n";
print $log_fh "$cmd\n";
if ($dry_run == false) {
@results = `$cmd`;
#
# Pull the depth of coverage from pstacks.
#
my @lines = grep(/Mean coverage depth is/, @results);
my ($depth) = ($lines[0] =~ /^ Mean coverage depth is (\d+\.?\d*); Std Dev: .+; Max: .+$/);
push(@depths_of_cov, [$sample->{'file'}, $depth]);
}
write_results(\@results, $log_fh);
$i++;
$sample_id++;
}
write_depths_of_cov(\@depths_of_cov, $log_fh);
#
# Generate catalog of RAD loci.
#
print STDERR "Generating catalog...\n";
my $file_paths = "";
foreach $sample (@parents, @samples) {
$file_paths .= "-s $out_path/$sample->{'file'} ";
}
$cmd = $exe_path . "cstacks -g -b $batch_id -o $out_path $file_paths " . join(" ", @_cstacks) . " 2>&1";
print STDERR " $cmd\n";
print $log_fh "$cmd\n\n";
if ($dry_run == false) {
open($pipe_fh, "$cmd |");
while (<$pipe_fh>) {
print $log_fh $_;
if ($_ =~ /failed/i) { print STDERR "Catalog construction failed.\n"; exit(1); }
}
close($pipe_fh);
}
#
# Match parents, progeny, or samples to the catalog.
#
$file_paths = "";
print STDERR "Matching samples to the catalog...\n";
foreach $sample (@parents, @progeny, @samples) {
$file_paths .= "-s $out_path/$sample->{'file'} ";
}
$cat_file = "batch_" . $batch_id;
$cmd = $exe_path . "sstacks -g -b $batch_id -c $out_path/$cat_file -o $out_path $file_paths " . join(" ", @_sstacks) . " 2>&1";
print STDERR " $cmd\n";
print $log_fh "$cmd\n\n";
if ($dry_run == false) {
open($pipe_fh, "$cmd |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
}
if ($data_type eq "map") {
#
# Generate a set of observed haplotypes and a set of markers and generic genotypes
#
printf(STDERR "Generating genotypes...\n");
$cmd = $exe_path . "genotypes -b $batch_id -P $out_path -r 1 -c -s " . join(" ", @_genotypes) . " 2>&1";
print STDERR "$cmd\n";
print $log_fh "$cmd\n";
if ($dry_run == 0) {
open($pipe_fh, "$cmd |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
}
} else {
printf(STDERR "Calculating population-level summary statistics\n");
$cmd = $exe_path . "populations -b $batch_id -P $out_path -s " . join(" ", @_populations) . " 2>&1";
print STDERR "$cmd\n";
print $log_fh "$cmd\n";
if ($dry_run == 0) {
open($pipe_fh, "$cmd |");
while (<$pipe_fh>) {
print $log_fh $_;
}
close($pipe_fh);
}
}
}
sub parse_population_map {
my ($sample_list, $pop_ids, $pops, $grp_ids, $grps) = @_;
my ($fh, @parts, $line, $sample);
return if (length($popmap_path) == 0);
open($fh, "<$popmap_path") or die("Unable to open population map, '$popmap_path', $!\n");
while ($line = <$fh>) {
chomp $line;
next if ($line =~ /^\s*#/);
@parts = split(/\t/, $line);
if (scalar(@parts) > 3) {
die("Unable to parse population map, '$popmap_path' (map should contain no more than three columns).\n");
}
push(@{$sample_list}, $parts[0]);
$pop_ids->{$parts[0]} = $parts[1];
$pops->{$parts[1]}++;
if (scalar(@parts) > 2) {
$grp_ids->{$parts[0]} = $parts[2];
$grps->{$parts[2]}++;
}
}
if (scalar(keys %{$grps}) == 0) {
$grps->{"1"}++;
foreach $sample (@{$sample_list}) {
$grp_ids->{$sample} = "1";
}
}
print STDERR "Parsed population map: ", scalar(@{$sample_list}), " files in ", scalar(keys %{$pops});
scalar(keys %{$pops}) == 1 ? print STDERR " population" : print STDERR " populations";
print STDERR " and ", scalar(keys %{$grps});
scalar(keys %{$grps}) == 1 ? print STDERR " group.\n" : print STDERR " groups.\n";
close($fh);
}
sub initialize_samples {
my ($parents, $progeny, $samples, $sample_list, $pop_ids, $grp_ids) = @_;
my ($local_gzip, $file, $prefix, $suffix, $path, $found, $i);
if (scalar(@{$sample_list}) > 0 && scalar(@{$samples}) == 0) {
my @suffixes = ("sam", "bam", "map", "bowtie");
my @fmts = ("sam", "bam", "map", "bowtie");
#
# If a population map was specified and no samples were provided on the command line.
#
foreach $sample (@{$sample_list}) {
$found = false;
for ($i = 0; $i < scalar(@suffixes); $i++) {
$path = $sample_path . $sample . "." . $suffixes[$i];
if (-e $path) {
if ($i == 1) {
$gzip = true;
}
push(@{$samples}, {'path' => $sample_path,
'file' => $sample,
'suffix' => $suffixes[$i],
'type' => "sample",
'fmt' => $fmts[$i]});
$found = true;
last;
}
}
if ($found == false) {
die("Unable to find sample '$sample' in directory '$sample_path' as specified in the population map, '$popmap_path'.\n");
}
}
} else {
#
# Process any samples were specified on the command line.
#
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
($prefix, $suffix) = ($sample->{'path'} =~ /^(.+)\.(.+)$/);
$sample->{'suffix'} = $suffix;
if ($prefix =~ /^.*\/.+$/) {
($path, $file) = ($prefix =~ /^(.*\/)(.+)$/);
} else {
$file = $prefix;
$path = "";
}
$sample->{'path'} = $path;
$sample->{'file'} = $file;
if ($suffix =~ /^bam$/) {
$sample->{'fmt'} = "bam";
$gzip = true;
} elsif ($suffix =~ /^sam$/) {
$sample->{'fmt'} = "sam";
} elsif ($suffix =~ /^map$/) {
$sample->{'fmt'} = "map";
} elsif ($suffix =~ /^bowtie$/) {
$sample->{'fmt'} = "bowtie";
} else {
die("Unknown input file type for file '" . $sample->{'path'} . "'.\n");
}
$path = $sample->{'path'} . $sample->{'file'} . "." . $sample->{'suffix'};
if (!-e $path) {
die("Unable to locate sample file '$path'\n");
}
}
foreach $sample (@{$parents}) {
$sample->{'type'} = "parent";
}
foreach $sample (@{$progeny}) {
$sample->{'type'} = "progeny";
}
foreach $sample (@{$samples}) {
$sample->{'type'} = "sample";
}
}
#
# If a population map was specified, make sure all samples in the list were found (and vice versa) and assign popualtion IDs.
#
if (scalar(@{$sample_list}) > 0) {
my %sample_hash;
foreach $sample (@{$samples}) {
$sample_hash{$sample->{'file'}}++;
if (!defined($pop_ids->{$sample->{'file'}})) {
die("Unable to find an entry for '" . $sample->{'file'} . "' in the population map, '$popmap_path'.\n");
} else {
$sample->{'pop_id'} = $pop_ids->{$sample->{'file'}};
}
if (!defined($grp_ids->{$sample->{'file'}})) {
die("Unable to find an entry for '" . $sample->{'file'} . "' in the population map, '$popmap_path'.\n");
} else {
$sample->{'grp_id'} = $grp_ids->{$sample->{'file'}};
}
}
foreach $sample (@{$sample_list}) {
if (!defined($sample_hash{$sample})) {
die("Unable to find a file corresponding to the population map entry '" . $sample . "' in the population map, '$popmap_path'.\n");
}
}
} else {
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
$sample->{'pop_id'} = "1";
$sample->{'grp_id'} = "1";
$pop_ids->{$sample->{'file'}} = $sample->{'pop_id'};
$grp_ids->{$sample->{'file'}} = $sample->{'grp_id'};
}
}
#
# Check that no duplicate files were specified.
#
my (%files, $file);
foreach $file (@{$parents}, @{$progeny}, @{$samples}) {
$files{$file}++;
}
foreach $file (keys %files) {
if ($files{$file} > 1) {
die("A duplicate file was specified which may create undefined results, '$file'\n");
}
}
print STDERR "Found ", scalar(@{$parents}), " parental file(s).\n" if (scalar(@{$parents}) > 0);
print STDERR "Found ", scalar(@{$progeny}), " progeny file(s).\n" if (scalar(@{$progeny}) > 0);
print STDERR "Found ", scalar(@{$samples}), " sample file(s).\n" if (scalar(@{$samples}) > 0);
}
sub initialize_database {
my ($log_fh, $parents, $progeny, $samples, $sample_ids) = @_;
my (@results, $sample_id, $sample);
print $log_fh "Initializing the database...\n";
#
# Create the database.
#
if ($create_db) {
#
# Check that the database doesn't already exist.
#
if ($dry_run == false) {
@results = `mysql --defaults-file=$cnf -N -B -e "SHOW DATABASES LIKE '$db'"`;
if (scalar(@results) > 0 && $overw_db == false) {
die("Unable to create database '$db', it already exists.\n");
}
}
if ($overw_db == true) {
`mysql --defaults-file=$cnf -N -B -e "DROP DATABASE IF EXISTS $db"` if ($dry_run == false);
print $log_fh "mysql --defaults-file=$cnf -N -B -e \"DROP DATABASE IF EXISTS '$db'\"\n";
}
`mysql --defaults-file=$cnf -e "CREATE DATABASE $db"` if ($dry_run == false);
print $log_fh "mysql --defaults-file=$cnf $db -e \"CREATE DATABASE $db\"\n";
`mysql --defaults-file=$cnf $db < $mysql_tables` if ($dry_run == false);
print $log_fh "mysql --defaults-file=$cnf $db < $mysql_tables\n";
}
#
# Set the SQL Batch ID for this set of loci, along with description and date of
# sequencing. Insert this batch data into the database.
#
`mysql --defaults-file=$cnf $db -e "INSERT INTO batches SET id=$batch_id, description='$desc', date='$date', type='$data_type'"` if ($dry_run == false);
print $log_fh "mysql --defaults-file=$cnf $db -e \"INSERT INTO batches SET id=$batch_id, description='$desc', date='$date', type='$data_type'\"\n";
print $log_fh "Loading sample data into the MySQL database...\n";
my $i = 1;
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
if ($dry_run == false) {
`mysql --defaults-file=$cnf $db -e "INSERT INTO samples SET sample_id=$i, batch_id=$batch_id, type='$sample->{'type'}', file='$sample->{'file'}', pop_id='$sample->{'pop_id'}', group_id='$sample->{'grp_id'}'"`;
@results = `mysql --defaults-file=$cnf $db -N -B -e "SELECT id FROM samples WHERE sample_id=$i AND batch_id=$batch_id AND type='$sample->{'type'}' AND file='$sample->{'file'}'"`;
chomp $results[0];
$sample_id = $results[0];
#
# Save the sample ID to use when running pstacks.
#
$sample_ids->{$sample->{'file'}} = $sample_id;
}
print $log_fh "mysql --defaults-file=$cnf $db -e \"INSERT INTO samples SET sample_id=$i, batch_id=$batch_id, type='$sample->{'type'}', file='$sample->{'file'}', pop_id='$sample->{'pop_id'}', group_id='$sample->{'grp_id'}'\"\n";
$i++;
}
print $log_fh "\n";
}
sub load_sql_data {
my ($log_fh, $pops, $parents, $progeny, $samples) = @_;
my ($pop_cnt, $sample, $num_files, $i, $file);
print STDERR "\nComputation is complete, loading results to the database '$db'.\n";
my $pop_cnt = scalar(keys %{$pops});
$i = 1;
$num_files = scalar(@{$parents}) + scalar(@{$progeny}) + scalar(@{$samples});
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
printf(STDERR "Loading pstacks output to $db; file % 3s of % 3s [%s]...", $i, $num_files, $sample->{'file'});
if ($gzip == true) {
$file = "$out_path/$sample->{'file'}" . ".tags.tsv.gz";
import_gzsql_file($log_fh, $file, "unique_tags", 1);
$file = "$out_path/$sample->{'file'}" . ".snps.tsv.gz";
import_gzsql_file($log_fh, $file, "snps", 1);
$file = "$out_path/$sample->{'file'}" . ".alleles.tsv.gz";
import_gzsql_file($log_fh, $file, "alleles", 1);
} else {
$file = "$out_path/$sample->{'file'}" . ".tags.tsv";
import_sql_file($log_fh, $file, "unique_tags", 1);
$file = "$out_path/$sample->{'file'}" . ".snps.tsv";
import_sql_file($log_fh, $file, "snps", 1);
$file = "$out_path/$sample->{'file'}" . ".alleles.tsv";
import_sql_file($log_fh, $file, "alleles", 1);
}
print STDERR "done.\n";
$i++;
}
print STDERR "Importing catalog to $db...";
my $cat_file = "batch_" . $batch_id;
if ($gzip == true) {
$file = "$out_path/$cat_file" . ".catalog.tags.tsv.gz";
import_gzsql_file($log_fh, $file, "catalog_tags", 1);
$file = "$out_path/$cat_file" . ".catalog.snps.tsv.gz";
import_gzsql_file($log_fh, $file, "catalog_snps", 1);
$file = "$out_path/$cat_file" . ".catalog.alleles.tsv.gz";
import_gzsql_file($log_fh, $file, "catalog_alleles", 1);
} else {
$file = "$out_path/$cat_file" . ".catalog.tags.tsv";
import_sql_file($log_fh, $file, "catalog_tags", 1);
$file = "$out_path/$cat_file" . ".catalog.snps.tsv";
import_sql_file($log_fh, $file, "catalog_snps", 1);
$file = "$out_path/$cat_file" . ".catalog.alleles.tsv";
import_sql_file($log_fh, $file, "catalog_alleles", 1);
}
print STDERR "done.\n";
#
# Load the sstacks results to the database if requested.
#
$i = 1;
foreach $sample (@{$parents}, @{$progeny}, @{$samples}) {
printf(STDERR "Loading sstacks output to $db; file % 3s of % 3s [%s]...", $i, $num_files, $sample->{'file'});
if ($gzip == true) {
$file = "$out_path/" . $sample->{'file'} . ".matches.tsv.gz";
import_gzsql_file($log_fh, $file, "matches", 1);
} else {
$file = "$out_path/" . $sample->{'file'} . ".matches.tsv";
import_sql_file($log_fh, $file, "matches", 1);
}
print STDERR "done.\n";
$i++;
}
if ($data_type eq "map") {
#
# Load the outputs generated by genotypes to the database.
#
$file = "$out_path/batch_" . $batch_id . ".markers.tsv";
import_sql_file($log_fh, $file, "markers", 1);
$file = "$out_path/batch_" . $batch_id . ".genotypes_1.txt";
import_sql_file($log_fh, $file, "catalog_genotypes", 1);
} else {
#
# Load the outputs generated by populations to the database.
#
$file = "$out_path/batch_" . $batch_id . ".markers.tsv";
import_sql_file($log_fh, $file, "markers", 1);
$file = "$out_path/batch_" . $batch_id . ".sumstats.tsv";
import_sql_file($log_fh, $file, "sumstats", $pop_cnt+1);
$file = "$out_path/batch_" . $batch_id . ".hapstats.tsv";
import_sql_file($log_fh, $file, "hapstats", $pop_cnt+1);
#
# Import the Fst files.
#
my $fst_cnt = 0;
my (@keys, $m, $n);
@keys = sort keys %pops;
for ($m = 0; $m < scalar(@keys); $m++) {
for ($n = 0; $n < scalar(@keys); $n++) {
$file = "$out_path/batch_" . $batch_id . ".fst_" . $keys[$m] . "-" . $keys[$n] . ".tsv";
if (-e $file) {
import_sql_file($log_fh, $file, "fst", 1);
$fst_cnt++;
}
}
}
print STDERR "Imported $fst_cnt Fst file(s).\n";
#
# Import the Phi_st files.
#
$fst_cnt = 0;
for ($m = 0; $m < scalar(@keys); $m++) {
for ($n = 0; $n < scalar(@keys); $n++) {
$file = "$out_path/batch_" . $batch_id . ".phistats_" . $keys[$m] . "-" . $keys[$n] . ".tsv";
if (-e $file) {
import_sql_file($log_fh, $file, "phist", 3);
$fst_cnt++;
}
}
}
print STDERR "Imported $fst_cnt Haplotype Fst file(s).\n";
}
print $log_fh "\n";
#
# Index the radtags database
#
my ($cmd, @results);
print STDERR "Indexing the database...\n";
$cmd = $exe_path . "index_radtags.pl -D $db -t -c 2>&1";
print STDERR "$cmd\n";
print $log_fh "$cmd\n";
@results = `$cmd` if ($dry_run == false);
print $log_fh @results;
}
sub write_results {
my ($results, $log_fh) = @_;
my $line;
foreach $line (@{$results}) {
if ($line =~ /\r/) {
$line =~ s/^.+\r(.*\n)$/\1/;
}
print $log_fh $line;
}
}
sub write_depths_of_cov {
my ($depths, $log_fh) = @_;
print STDERR "\nDepths of Coverage for Processed Samples:\n";
print $log_fh "\nDepths of Coverage for Processed Samples:\n";
foreach $a (@{$depths}) {
print STDERR $a->[0], ": ", $a->[1], "x\n";
print $log_fh $a->[0], ": ", $a->[1], "x\n";
}
print STDERR "\n";
print $log_fh "\n";
}
sub import_sql_file {
my ($log_fh, $file, $table, $skip_lines) = @_;
my (@results, $ignore);
$ignore = "IGNORE $skip_lines LINES" if ($skip_lines > 0);
@results = `mysql --defaults-file=$cnf $db -e "LOAD DATA LOCAL INFILE '$file' INTO TABLE $table $ignore"` if ($sql == true && $dry_run == false);
if ($sql == true) {
print $log_fh "mysql --defaults-file=$cnf $db -e \"LOAD DATA LOCAL INFILE '$file' INTO TABLE $table $ignore\"\n", @results;
}
}
sub import_gzsql_file {
my ($log_fh, $file, $table, $skip_lines) = @_;
my (@results, $ignore);
$ignore = "IGNORE $skip_lines LINES" if ($skip_lines > 0);
#
# Get a temporary file name and create a named pipe.
#
my $tmpdir = File::Spec->tmpdir();
my $named_pipe = mktemp($tmpdir . "/denovo_map_XXXXXX");
if ($sql == true && $dry_run == false) {
mkfifo($named_pipe, 0700) || die("Unable to create named pipe for loading gzipped data: $named_pipe, $!");
print $log_fh "Streaming $file into named pipe $named_pipe.\n";
}
#
# Dump our gzipped data onto the named pipe.
#
system("gunzip -c $file > $named_pipe &") if ($sql == true && $dry_run == false);
@results = `mysql --defaults-file=$cnf $db -e "LOAD DATA LOCAL INFILE '$named_pipe' INTO TABLE $table $ignore"` if ($sql == true && $dry_run == false);
if ($sql == 1) {
print $log_fh "mysql --defaults-file=$cnf $db -e \"LOAD DATA LOCAL INFILE '$named_pipe' INTO TABLE $table $ignore\"\n", @results;
}
#
# Remove the pipe.
#
unlink($named_pipe) if ($sql == true && $dry_run == false);
}
sub parse_command_line {
my ($arg);
while (@ARGV) {
$_ = shift @ARGV;
if ($_ =~ /^-v$/) { version(); exit(); }
elsif ($_ =~ /^-h$/) { usage(); }
elsif ($_ =~ /^-p$/) { push(@parents, { 'path' => shift @ARGV }); }
elsif ($_ =~ /^-r$/) { push(@progeny, { 'path' => shift @ARGV }); }
elsif ($_ =~ /^-s$/) { push(@samples, { 'path' => shift @ARGV }); }
elsif ($_ =~ /^-d$/) { $dry_run = true; }
elsif ($_ =~ /^-o$/) { $out_path = shift @ARGV; }
elsif ($_ =~ /^-D$/) { $desc = shift @ARGV; }
elsif ($_ =~ /^-e$/) { $exe_path = shift @ARGV; }
elsif ($_ =~ /^-b$/) { $batch_id = shift @ARGV; }
elsif ($_ =~ /^-i$/) { $sample_id = shift @ARGV; }
elsif ($_ =~ /^-a$/) { $date = shift @ARGV; }
elsif ($_ =~ /^-S$/) { $sql = false; }
elsif ($_ =~ /^-B$/) { $db = shift @ARGV; }
elsif ($_ =~ /^-m$/) { $min_cov = shift @ARGV; }
elsif ($_ =~ /^-P$/) { $min_rcov = shift @ARGV; }
elsif ($_ =~ /^--samples$/) {
$sample_path = shift @ARGV;
} elsif ($_ =~ /^-O$/) {
$popmap_path = shift @ARGV;
push(@_populations, "-M " . $popmap_path);
} elsif ($_ =~ /^--create_db$/) {
$create_db = true;
} elsif ($_ =~ /^--overw_db$/) {
$overw_db = true;
$create_db = true;
} elsif ($_ =~ /^-A$/) {
$arg = shift @ARGV;
push(@_genotypes, "-t " . $arg);
$arg = lc($arg);
if ($arg ne "gen" && $arg ne "cp" && $arg ne "f2" && $arg ne "bc1" && $arg ne "dh") {
print STDERR "Unknown genetic mapping cross specified: '$arg'\n";
usage();
}
} elsif ($_ =~ /^-T$/) {
$arg = shift @ARGV;
push(@_pstacks, "-p " . $arg);
push(@_cstacks, "-p " . $arg);
push(@_sstacks, "-p " . $arg);
push(@_populations, "-t " . $arg);
} elsif ($_ =~ /^--bound_low$/) {
push(@_pstacks, "--bound_low " . shift @ARGV);
push(@_pstacks, "--model_type bounded");
} elsif ($_ =~ /^--bound_high$/) {
push(@_pstacks, "--bound_high " . shift @ARGV);
push(@_pstacks, "--model_type bounded");
} elsif ($_ =~ /^--alpha$/) {
push(@_pstacks, "--alpha " . shift @ARGV);
} elsif ($_ =~ /^-X$/) {
#
# Pass an arbitrary command-line option to a pipeline program.
#
# Command line option must be of the form '-X "program:option"'
#
$arg = shift @ARGV;
my ($prog, $opt) = ($arg =~ /^(\w+):(.+)$/);
if ($prog eq "pstacks") {
push(@_pstacks, $opt);
} elsif ($prog eq "cstacks") {
push(@_cstacks, $opt);
} elsif ($prog eq "sstacks") {
push(@_sstacks, $opt);
} elsif ($prog eq "genotypes") {
push(@_genotypes, $opt);
} elsif ($prog eq "populations") {
push(@_populations, $opt);
} else {
print STDERR "Unknown pipeline program, '$arg'\n";
usage();
}
}
else {
print STDERR "Unknown command line option: '$_'\n";
usage();
}
}
$exe_path = $exe_path . "/" if (substr($exe_path, -1) ne "/");
$out_path = substr($out_path, 0, -1) if (substr($out_path, -1) eq "/");
if ($batch_id !~ /^\d+$/ || $batch_id < 0) {
print STDERR "You must specify a batch ID and it must be an integer (e.g. 1, 2, 3).\n";
usage();
}
if ($sql == true && length($date) == 0) {
$date = strftime("%Y-%m-%d", (localtime(time)));
}
if (scalar(@parents) > 0 && scalar(@samples) > 0) {
print STDERR "You must specify either parent or sample files, but not both.\n";
usage();
}
if (scalar(@parents) == 0 && scalar(@samples) == 0 && length($popmap_path) == 0) {
print STDERR "You must specify at least one parent or sample file.\n";
usage();
}
if (length($sample_path) > 0) {
$sample_path .= "/" if (substr($sample_path, -1) ne "/");
}
if (scalar(@samples) > 0 || length($popmap_path) > 0) {
$data_type = "population";
} else {
$data_type = "map";
}
}
sub version {
print STDERR "ref_map.pl ", stacks_version, "\n";
}
sub usage {
version();
print STDERR <<EOQ;
ref_map.pl -p path -r path [-s path] -o path [-m min_cov] [-T num_threads] [-A type] [-O popmap] [-B db -b batch_id -D "desc"] [-S -i num] [-e path] [-d] [-h]
b: batch ID representing this dataset (an integer, e.g. 1, 2, 3).
o: path to write pipeline output files.
O: if analyzing one or more populations, specify a pOpulation map.
A: if processing a genetic map, specify the cross type, 'CP', 'F2', 'BC1', 'DH', or 'GEN'.
T: specify the number of threads to execute.
e: executable path, location of pipeline programs.
d: perform a dry run. Do not actually execute any programs, just print what would be executed.
h: display this help message.
Specify each sample separately:
p: path to a Bowtie/SAM/BAM file containing one set of parent sequences from a mapping cross.
r: path to a Bowtie/SAM/BAM file containing one set of progeny sequences from a mapping cross.
s: path to a Bowtie/SAM/BAM file containing an individual sample from a population.
Specify a path to samples and provide a population map:
--samples <path>: specify a path to the directory of samples (samples will be read from population map).
Stack assembly options:
m: specify a minimum number of identical, raw reads required to create a stack (default 3).
P: specify a minimum number of identical, raw reads required to create a stack in 'progeny' individuals.
Database options:
B: specify a database to load data into.
D: a description of this batch to be stored in the database.
S: disable recording SQL data in the database.
i: starting sample_id, this is determined automatically if database interaction is enabled.
--create_db: create the database specified by '-B' and populate the tables.
--overw_db: delete the database before creating a new copy of it (turns on --create_db).
SNP Model Options (these options are passed on to pstacks):
--bound_low <num>: lower bound for epsilon, the error rate, between 0 and 1.0 (default 0).
--bound_high <num>: upper bound for epsilon, the error rate, between 0 and 1.0 (default 1).
--alpha <num>: chi square significance level required to call a heterozygote or homozygote, either 0.1, 0.05 (default), 0.01, or 0.001.
Arbitrary command line options:
-X "program:option": pass a command line option to one of the pipeline components, e.g.'-X "sstacks:-x"'.
EOQ
exit(0);
}
|