=head1 LICENSE Copyright [1999-2016] Wellcome Trust Sanger Institute and the EMBL-European Bioinformatics Institute Licensed under the Apache License, Version 2.0 (the "License"); you may not use this file except in compliance with the License. You may obtain a copy of the License at http://www.apache.org/licenses/LICENSE-2.0 Unless required by applicable law or agreed to in writing, software distributed under the License is distributed on an "AS IS" BASIS, WITHOUT WARRANTIES OR CONDITIONS OF ANY KIND, either express or implied. See the License for the specific language governing permissions and limitations under the License. =cut package AssemblyMapper::BlastzAligner; =head1 NAME BlastzAligner.pm - module to do a whole genome alignment between two closely related assemblies and create assembly entries from it. =head1 SYNOPSIS my $support = new Bio::EnsEMBL::Utils::ConversionSupport($SERVERROOT); my $aligner = BlastzAligner->new(-SUPPORT => $support); # create a tempdir for storing input and output $aligner->create_tempdir; # write sequences to fasta and nib files $aligner->write_sequence( $alt_slice, $support->param('altassembly'), "alt_sequence.1" ); $aligner->write_sequence( $ref_slice, $support->param('assembly'), "ref_sequence.1" ); # run blastz $aligner->run_blastz("alt_sequence.1", "ref_sequence.1"); =head1 DESCRIPTION This modules contains helper functions to generate a whole genome alignment between two closely related assemblies using blastz from scratch. Alignments are then stored in an Ensembl assembly table. The module is part of a series of scripts to create a mapping between two assemblies. See "Related scripts" below for an overview of the whole process. =head1 RELATED SCRIPTS The whole process of creating a whole genome alignment is done by these scripts: ensembl/misc-scripts/assembly/load_alternative_assembly.pl ensembl/misc-scripts/assembly/align_by_clone_identity.pl ensembl/misc-scripts/assembly/align_nonident_regions.pl See documention in the respective script for more information. =head1 AUTHOR Patrick Meidl , Ensembl core API team =head1 CONTACT Please post comments/questions to the Ensembl development list =cut use strict; use warnings; no warnings 'uninitialized'; use Bio::EnsEMBL::Utils::Argument qw(rearrange); use File::Path; use constant FMT1 => "%-30s%10.0f (%3.2f%%)\n"; use constant FMT2 => "%-30s%10.0f\n"; use constant FMT3 => "%-8s%-12s%-5s%-10s%-10s%-10s%-10s\n"; use constant FMT4 => "%-8s%-12s%-5s%8.0f %8.0f %8.0f %8.0f\n"; =head2 new Arg[-SUPPORT] : a Bio::EnsEMBL::Utils::ConversionSupport object Example : my $support = new Bio::EnsEMBL::Utils::ConversionSupport( $SERVERROOT); my $aligner = BlastzAligner->new(-SUPPORT => $support); Description : object constructor method Return type : a BlastzAligner object Exceptions : none Caller : general =cut sub new { my ($class, @args) = @_; my $self = {}; bless $self, $class; my ($support) = rearrange([qw(SUPPORT)], @args); $self->support($support); # set bindir $self->bindir($self->support->param('bindir') || '/usr/local/ensembl/bin'); return $self; } =head2 create_tempdir Arg[1] : String $tmpdir - temporary directory name Example : $aligner->create_tempdir; Description : Creates a temporary directory in /tmp with a semi-random name (username.timestamp.randomnumber). Alternatively, you can pass it the name of a directory to use. Return type : String - name of the tempdir created Exceptions : Thrown if tempdir can't be created Caller : general =cut sub create_tempdir { my $self = shift; my $tempdir = shift; if ($tempdir) { unless (-d $tempdir) { $self->support->log_error("Can't find tempdir $tempdir: $!"); } } else { # create tmpdir to store input and output my $user = `whoami`; chomp $user; $tempdir = "/tmp/$user.".time.".".int(rand(100000)); $self->support->log("Creating tmpdir $tempdir...\n"); system("mkdir $tempdir") == 0 or $self->support->log_error("Can't create tmp dir $tempdir: $!\n"); $self->support->log("Done.\n"); } $self->tempdir($tempdir); return $tempdir; } =head2 remove_tempdir Example : $aligner->remove_tempdir; Description : Removes a temporary directory created by $self->create_tempdir. Return type : none Exceptions : none Caller : general =cut sub remove_tempdir { my $self = shift; rmtree($self->tempdir) or $self->support->log_warning("Could not delete ".$self->tempdir.": $!\n"); } =head2 write_sequence Arg[1] : Bio::EnsEMBL::Slice $slice - slice for which to write sequence Arg[2] : String $assembly - assembly name Arg[3] : String $basename1 - basename of single sequence file Arg[4] : (optional) String $basename2 - basename of multiple sequence file Example : $aligner->write_sequence($slice, 'NCBI35', 'e_seq.1'); Description : Writes a slice's sequence to a fasta file and converts it to nib format. Optionally appends the sequence to another multi-sequence fasta file. Return type : none Exceptions : thrown if faToNib or file appending fails Caller : general =cut sub write_sequence { my ($self, $slice, $assembly, $basename1, $basename2) = @_; my $tmpdir = $self->tempdir; unless (-e "$tmpdir/$basename1.fa") { my $fh = $self->support->filehandle('>', "$tmpdir/$basename1.fa"); my $cs_name = $slice->coord_system_name; print $fh join(':', ">$basename1 dna:chromfrag $cs_name", $assembly, $slice->start, $slice->end, $slice->strand ), "\n"; print $fh $slice->get_repeatmasked_seq(undef, 1)->seq, "\n"; close($fh); } # convert fasta to nib (needed for lavToAxt) unless (-e "$tmpdir/$basename1.nib") { system($self->bindir."/faToNib $tmpdir/$basename1.fa $tmpdir/$basename1.nib") == 0 or $self->support->log_error("Can't run faToNib: $!\n"); } if ($basename2) { system("cat $tmpdir/$basename1.fa >> $tmpdir/$basename2.fa") == 0 or $self->support->log_error("Can't concat fasta files: $!\n"); } } =head2 run_blastz Arg[1] : String $A_basename - basename of alternative fasta file Arg[2] : String $R_basename - basename of reference fasta file Example : $aligner->run_blastz('alt_seq.1', 'ref_seq.1'); Description : Runs blastz between an alternative and multiple reference sequences. Return type : none Exceptions : thrown if blastz fails Caller : general =cut sub run_blastz { my ($self, $A_basename, $R_basename) = @_; my $tmpdir = $self->tempdir; my $bindir = $self->bindir; my $id = $self->id; my $blastz_cmd = qq($bindir/blastz $tmpdir/$A_basename.fa $tmpdir/$R_basename.fa Q=blastz_matrix.txt T=0 L=10000 H=2200 Y=3400 > $tmpdir/blastz.$id.lav); unless (-e "$tmpdir/blastz.$id.lav") { system($blastz_cmd) == 0 or $self->support->log_error("Can't run blastz: $!\n"); } } =head2 lav_to_axt Example : $aligner->lav_to_axt; Description : Converts blastz output from lav to axt format. Target and query sequences must be present in nib format in the temporary directory for this to work. Return type : none Exceptions : thrown if lavToAxt fails Caller : general =cut sub lav_to_axt { my $self = shift; my $tmpdir = $self->tempdir; my $id = $self->id; unless (-e "$tmpdir/blastz.$id.axt") { system($self->bindir."/lavToAxt $tmpdir/blastz.$id.lav $tmpdir $tmpdir $tmpdir/blastz.$id.axt") == 0 or $self->support->log_error("Can't run lavToAxt: $!\n"); } } =head2 find_best_alignment Example : $aligner->find_best_alignment; Description : Finds the best set of non-overlapping alignments by running axtBest. Return type : none Exceptions : thrown if axtBest fails Caller : general =cut sub find_best_alignment { my $self = shift; my $tmpdir = $self->tempdir; my $id = $self->id; unless (-e "$tmpdir/blastz.$id.best.axt") { system($self->bindir."/axtBest $tmpdir/blastz.$id.axt all $tmpdir/blastz.$id.best.axt") == 0 or $self->support->log_error("Can't run axtBest: $!\n"); } } =head2 parse_blastz_output Example : $aligner->parse_blastz_output; Description : Reads a blastz alignment result from an axt file and creates a datastructure containing ungapped alignments from it. Note that mismatches are allowed, but separate stats will be collected for them. Return type : none Exceptions : none Caller : general =cut sub parse_blastz_output { my $self = shift; # read file my $tmpdir = $self->tempdir; my $id = $self->id; my $fh = $self->support->filehandle('<', "$tmpdir/blastz.$id.best.axt"); # initialize stats $self->init_stats(qw(match mismatch gap alignments bp)); my $i = 1; my ($header, $A_seq, $R_seq); while (my $line = <$fh>) { # there are blocks of 4 lines, where line 1 is the header, line 2 is # A_seq, line3 is R_seq $header = $line unless (($i-1) % 4); $A_seq = $line unless (($i-2) % 4); chomp $A_seq; my @A_arr = split(//, $A_seq); $R_seq = $line unless (($i-3) % 4); chomp $R_seq; my @R_arr = split(//, $R_seq); # compare sequences letter by letter if ($i % 4 == 0) { my $match_flag = 0; $self->init_stats(qw(A_gap R_gap)); my %coords; @coords{'R_id', 'A_start', 'A_end', 'R_start', 'R_end', 'strand'} = (split(/ /, $header))[4, 2, 3, 5, 6, 7]; $coords{'R_id'} =~ s/ref_seq\.(.*)/$1/; ($coords{'strand'} eq '+') ? ($coords{'strand'} = 1) : ($coords{'strand'} = -1); for (my $j = 0; $j < scalar(@A_arr); $j++) { # gap if ($A_arr[$j] eq '-' or $R_arr[$j] eq '-') { $self->stats_incr('gap', 1); $self->stats_incr('A_gap', 1) if ($A_arr[$j] eq '-'); $self->stats_incr('R_gap', 1) if ($R_arr[$j] eq '-'); $match_flag = 0; } else { $self->found_match($match_flag, $j, \%coords); $match_flag = 1; # match if ($A_arr[$j] eq $R_arr[$j]) { $self->stats_incr('match', 1); # mismatch } else { $self->stats_incr('mismatch', 1); } } } $self->stats_incr('bp', scalar(@A_arr)); $self->stats_incr('alignments', 1); } $i++; } } =head2 cleanup_tmpfiles Arg[1-N] : (optional) list @files - additional tmp files to delete Example : $self->cleanup_tmpfiles('e_seq.fa', 'v_seq.fa'); Description : deletes temporary files Return type : none Exceptions : Warning if file cannot be deleted Caller : general Status : stable =cut sub cleanup_tmpfiles { my $self = shift; my @files = @_; my $tmpdir = $self->tempdir; my $id = $self->id; push @files, "blastz.$id.lav", "blastz.$id.axt", "blastz.$id.best.axt"; foreach my $file (@files) { unlink("$tmpdir/$file") or $self->support->log_warning("Couldn't delete file $file: $!"); } } =head2 found_match Arg[1] : Boolean $match_flag - flag indicating if last bp was a match Arg[2] : Int $j - current bp position in the alignment Arg[3] : Hashref $coords - alignment coordinates and strand from blastz output Description : Populates a datastructure describing blocks of alignment. Return type : none Exceptions : none Caller : internal =cut sub found_match { my ($self, $match_flag, $j, $coords) = @_; my $id = $self->id; my $align = $self->get_stats('alignments'); my $R_chr = $self->seq_region_name; # last position was a match if ($match_flag) { # adjust align block end if ($self->{'_match'}->{$R_chr}->{$id}->[$align]) { my $c = scalar(@{ $self->{'_match'}->{$R_chr}->{$id}->[$align] }) - 1; $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1] = $coords->{'A_start'} + $j - $self->get_stats('A_gap'); $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3] = $coords->{'R_start'} + $j - $self->get_stats('R_gap'); } # last position was a non-match } else { # start a new align block push @{ $self->{'_match'}->{$R_chr}->{$id}->[$align] }, [ $coords->{'A_start'} + $j - $self->get_stats('A_gap'), $coords->{'A_start'} + $j - $self->get_stats('A_gap'), $coords->{'R_start'} + $j - $self->get_stats('R_gap'), $coords->{'R_start'} + $j - $self->get_stats('R_gap'), $coords->{'strand'}, $coords->{'R_id'}, ]; } } =head2 adjust_coords Arg[1] : Int $A_start - start of alternatvie block in chromosomal coords Arg[2] : Int $A_end - end of alternative block in chromosomal coords Arg[3] : Arrayref $R_coords - list of start/end pairs of reference blocks in chromosomal coords Example : my $R_coords = [ [1, 1000], [3000, 5000] ]; $aligner->adjust_coords(1, 30000000, $R_coords); Description : Adjusts coordinates of blastz alignments to chromosomal coords. Return type : none Exceptions : none Caller : general =cut sub adjust_coords { my ($self, $A_start, $A_end, $R_coords) = @_; my $R_chr = $self->seq_region_name; my $id = $self->id; for (my $align = 0; $align < scalar(@{ $self->{'_match'}->{$R_chr}->{$id} || [] }); $align++) { for (my $c = 0; $c < scalar(@{ $self->{'_match'}->{$R_chr}->{$id}->[$align] || []}); $c++) { $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[0] += $A_start - 1; $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1] += $A_start - 1; # forward strand match if ($self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[4] == 1) { $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2] += $R_coords->{$self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[5]}->[0] - 1; $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3] += $R_coords->{$self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[5]}->[0] - 1; # reverse strand match } else { my $tmp_start = $R_coords->{$self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[5]}->[1] - $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3] + 1; $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3] = $R_coords->{$self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[5]}->[1] - $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2] + 1; $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2] = $tmp_start; } # sanity check: aligned region pairs must have same length my $A_len = $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1] - $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[0]; my $R_len = $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3] - $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2]; $self->support->log_warning("Length mismatch: $A_len <> $R_len in block $id, alignment $align, stretch $c\n", 2) unless ($A_len == $R_len); } } } =head2 filter_overlaps Description : DEPRECATED. This filtering algorithm didn't work well. Please run the separate script fix_overlaps.pl. =cut sub filter_overlaps { my $self = shift; my $id = $self->id; foreach my $R_chr (sort keys %{ $self->{'_match'} }) { my $coord_check = []; # rearrange the datastructure so that we can find overlaps foreach my $id (keys %{ $self->{'_match'}->{$R_chr} }) { for (my $align = 0; $align < scalar(@{ $self->{'_match'}->{$R_chr}->{$id} }); $align++) { for (my $c = 0; $c < scalar(@{ $self->{'_match'}->{$R_chr}->{$id}->[$align] || []}); $c++) { push @{ $coord_check }, [ $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[0], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3], $id, $align, $c, ]; } } } my @A_sort = sort { $a->[0] <=> $b->[0] } @{ $coord_check }; my @R_sort = sort { $a->[2] <=> $b->[2] } @{ $coord_check }; # sanity check: alternative alignments must not overlap (axtBest should # guarantee that) my $last; foreach my $c (@A_sort) { $self->support->log_warning("Overlapping alternative alignment at ".join(':', $R_chr, $c->[0], $c->[1])." (last_end ".$last->[1].")\n", 1) if ($last and $c->[0] <= $last->[1]); $last = $c; } # now filter reference overlaps my @seen; $last = undef; foreach my $c (@R_sort) { if ($last and $c->[2] <= $last->[3]) { $self->support->log_verbose("Overlapping reference alignment at ".join(':', $R_chr, $c->[2], $c->[3])." (last_end ".$last->[3].")\n", 1); # if last alignment was longer, delete this one if ($last->[3]-$last->[2] > $c->[3]-$c->[2]) { undef $self->{'_match'}->{$R_chr}->{$c->[4]}->[$c->[5]]->[$c->[6]]; # if last alignment was shorter, trace back and delete all # overlapping shorter alignments } else { foreach my $s (@seen) { # earlier alignment still overlapping if ($c->[2] <= $s->[3]) { # earlier alignment shorter -> delete it if ($s->[3]-$s->[2] < $c->[3]-$c->[2]) { undef $self->{'_match'}->{$R_chr}->{$s->[4]}->[$s->[5]]->[$s->[6]]; # this alignment shorter -> delete it } else { undef $self->{'_match'}->{$R_chr}->{$c->[4]}->[$c->[5]]->[$c->[6]]; $last = $s; last; } } else { $last = $s; last; } } $last = $c; } } unshift @seen, $c; $last = $c unless ($last); } } } =head2 write_assembly Arg[1] : Bio::*::DBSQL::DBAdaptor $R_dba - reference database adaptor Example : my $R_dba = $support->get_database('ensembl'); $aligner->write_assembly($R_dba); Description : Writes assembly entries for blastz alignments to the database. Return type : none Exceptions : none Caller : general =cut sub write_assembly { my ($self, $R_dba) = @_; my $R_dbh = $R_dba->dbc->db_handle; my $R_sa = $R_dba->get_SliceAdaptor; my $sth = $R_dbh->prepare(qq( INSERT IGNORE INTO assembly (asm_seq_region_id, cmp_seq_region_id, asm_start,asm_end, cmp_start, cmp_end, ori) VALUES (?, ?, ?, ?, ?, ?, ?) )); $self->support->log("Adding assembly entries for alignments...\n"); my $i; foreach my $R_chr (sort _by_chr_num keys %{ $self->{'_match'} }) { # get seq_region_id for alternative and reference chromosome my $A_chr = $R_chr; my $R_slice = $R_sa->fetch_by_region('toplevel', $R_chr, undef, undef, undef, $self->support->param('assembly')); my $R_sid = $R_sa->get_seq_region_id($R_slice); # we need to fetch the alternative slice from the reference db # explicitely by coord_system, since toplevel attribute is not set # there my $cs_name = $R_slice->coord_system_name; my $A_sid = $R_sa->get_seq_region_id($R_sa->fetch_by_region($cs_name, $A_chr, undef, undef, undef, $self->support->param('altassembly'))); foreach my $id (sort { $a <=> $b } keys %{ $self->{'_match'}->{$R_chr} }) { for (my $align = 0; $align < scalar(@{ $self->{'_match'}->{$R_chr}->{$id} }); $align++) { for (my $c = 0; $c < scalar(@{ $self->{'_match'}->{$R_chr}->{$id}->[$align] }); $c++) { if ($self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]) { $sth->execute( $R_sid, $A_sid, $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[2], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[3], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[0], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1], $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[4], ); $i++; } } } } } $self->support->log("Done inserting $i entries.\n"); } =head2 stats_incr Arg[1] : String $code - stats code Arg[2] : Int $incr - number by which stats should be incremented Example : $aligner->stats_incr('total_alignments', 1); Description : Increments stats. Return type : Int - stat value after increment Exceptions : none Caller : general =cut sub stats_incr { my ($self, $code, $incr) = @_; $self->{'_stats'}->{$code} += $incr; return $self->{'_stats'}->{$code}; } =head2 init_stats Arg[1] : Array @codes - stats codes to initialise Example : $aligner->init_stats('match', 'mismatch', 'alignments'); Description : Initialises stats (i.e. sets to 0). Return type : none Exceptions : none Caller : general =cut sub init_stats { my ($self, @codes) = @_; foreach my $code (@codes) { $self->{'_stats'}->{$code} = 0; } } =head2 get_stats Arg[1] : String $code - stats code Example : my $num_mismatches = $aligner->get_stats('mismatch'); Description : Stats getter. Return type : Int - stats value Exceptions : none Caller : general =cut sub get_stats { my ($self, $code) = @_; return $self->{'_stats'}->{$code}; } =head2 log_block_stats Arg[1] : Int $indent - indentation level Example : $aligner->log_block_stats(3); Description : Logs stats for an alignment block. Return type : none Exceptions : none Caller : general =cut sub log_block_stats { my ($self, $indent) = @_; $self->support->log("Blastz alignment stats:\n", $indent); $self->support->log(sprintf(FMT2, "Alignments:", $self->get_stats('alignments')), $indent+1); if ($self->get_stats('alignments')) { $self->support->log(sprintf(FMT1, "Matches:", $self->get_stats('match'), $self->get_stats('match')/$self->get_stats('bp')*100), $indent+1); $self->support->log(sprintf(FMT1, "Mismatches:", $self->get_stats('mismatch'), $self->get_stats('mismatch')/$self->get_stats('bp')*100), $indent+1); $self->support->log(sprintf(FMT1, "Gaps:", $self->get_stats('gap'), $self->get_stats('gap')/$self->get_stats('bp')*100), $indent+1); } map { $self->stats_incr($_.'_total', $self->get_stats($_)) } qw(match mismatch gap bp); } =head2 log_overall_stats Example : $aligner->log_overall_stats; Description : Logs overall alignment stats. Return type : none Exceptions : none Caller : general =cut sub log_overall_stats { my $self = shift; # blastz $self->support->log("\nOverall blastz alignment stats:\n"); unless ($self->get_stats('alignments')) { $self->support->log("No alignments found.\n", 1); return; } $self->support->log(sprintf(FMT1, "Matches:", $self->get_stats('match_total'), $self->get_stats('match_total')/$self->get_stats('bp_total')*100), 1); $self->support->log(sprintf(FMT1, "Mismatches:", $self->get_stats('mismatch_total'), $self->get_stats('mismatch_total')/$self->get_stats('bp_total')*100), 1); $self->support->log(sprintf(FMT1, "Gaps:", $self->get_stats('gap_total'), $self->get_stats('gap_total')/$self->get_stats('bp_total')*100), 1); # alignments to be written to assembly table $self->support->log_verbose("\nAlignments that will be written to assembly table:\n"); $self->support->log_verbose(sprintf(FMT3, qw(CHR BLOCK ALIGNMENT ALT_START ALT_END REF_START REF_END)), 1); $self->support->log_verbose(('-'x63)."\n", 1); foreach my $R_chr (sort _by_chr_num keys %{ $self->{'_match'} }) { foreach my $id (sort { $a <=> $b } keys %{ $self->{'_match'}->{$R_chr} }) { for (my $align = 0; $align < scalar(@{ $self->{'_match'}->{$R_chr}->{$id} }); $align++) { for (my $c = 0; $c < scalar(@{ $self->{'_match'}->{$R_chr}->{$id}->[$align] }); $c++) { if ($self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]) { $self->support->log_verbose(sprintf(FMT4, $R_chr, $id, $align+1, @{ $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c] }), 1); my $l = $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[1] - $self->{'_match'}->{$R_chr}->{$id}->[$align]->[$c]->[0]; $self->stats_incr('alignments_total', 1); $self->stats_incr('short1_10_total', 1) if ($l < 11); $self->stats_incr('short11_100_total', 1) if ($l > 10 and $l < 101); } } } } } $self->support->log("\nAssembly entry stats:\n"); $self->support->log(sprintf(FMT2, "Total alignment blocks:", $self->get_stats('alignments_total')), 1); $self->support->log(sprintf(FMT2, "Alignments 1-10 bp:", $self->get_stats('short1_10_total')), 1); $self->support->log(sprintf(FMT2, "Alignments 11-100 bp:", $self->get_stats('short11_100_total')), 1); } =head2 _by_chr_num Example : my @sorted = sort _by_chr_num qw(X, 6-COX, 14, 7); Description : Subroutine to use in sort for sorting chromosomes. Sorts numerically, then alphabetically Return type : values to be used by sort Exceptions : none Caller : internal =cut sub _by_chr_num { my @awords = split /-/, $a; my @bwords = split /-/, $b; my $anum = $awords[0]; my $bnum = $bwords[0]; if ($anum !~ /^[0-9]*$/) { if ($bnum !~ /^[0-9]*$/) { return $anum cmp $bnum; } else { return 1; } } if ($bnum !~ /^[0-9]*$/) { return -1; } if ($anum <=> $bnum) { return $anum <=> $bnum; } else { if ($#awords == 0) { return -1; } elsif ($#bwords == 0) { return 1; } else { return $awords[1] cmp $bwords[1]; } } } =head2 AUTOLOAD Arg[1] : (optional) String/Object - attribute to set Example : # setting a attribute $self->attr($val); # getting the attribute $self->attr; # undefining an attribute $self->attr(undef); Description : lazy function generator for getters/setters Return type : String/Object Exceptions : none Caller : general =cut sub AUTOLOAD { my $self = shift; my $attr = our $AUTOLOAD; $attr =~ s/.*:://; return unless $attr =~ /[^A-Z]/; no strict 'refs'; *{$AUTOLOAD} = sub { $_[0]->{'_data'}->{$attr} = $_[1] if (@_ > 1); return $_[0]->{'_data'}->{$attr}; }; $self->{'_data'}->{$attr} = shift if (@_); return $self->{'_data'}->{$attr}; } 1;