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# Bio::Tools::Alignment::Consed |
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# |
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# Please direct questions and support issues to |
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# |
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# Cared for by Chad Matsalla |
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# |
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# Copyright Chad Matsalla |
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# |
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# You may distribute this module under the same terms as perl itself |
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# POD documentation - main docs before the code |
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=head1 NAME |
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Bio::Tools::Alignment::Consed - A module to work with objects from consed .ace files |
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=head1 SYNOPSIS |
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# a report for sequencing stuff |
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my $o_consed = Bio::Tools::Alignment::Consed->new( |
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-acefile => "/path/to/an/acefile.ace.1", |
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-verbose => 1); |
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my $foo = $o_consed->set_reverse_designator("r"); |
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my $bar = $o_consed->set_forward_designator("f"); |
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# get the contig numbers |
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my @keys = $o_consed->get_contigs(); |
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# construct the doublets |
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my $setter_doublets = $o_consed->choose_doublets(); |
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# get the doublets |
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my @doublets = $o_consed->get_doublets(); |
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=head1 DESCRIPTION |
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L provides methods and objects to deal |
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with the output from the Consed software suite. Specifically, |
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takes an C<.ace> file and provides objects for the results. |
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A word about doublets: This module was written to accommodate a large |
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EST sequencing operation. In this case, EST's were sequenced from the |
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3' and from the 5' end of the EST. The objective was to find a |
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consensus sequence for these two reads. Thus, a contig of two is what |
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we wanted, and this contig should consist of the forward and reverse |
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reads of a getn clone. For example, for a forward designator of "F" |
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and a reverse designator of "R", if the two reads chad1F and chad1R |
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were in a single contig (for example Contig 5) it will be determined |
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that the consensus sequence for Contig 5 will be the sequence for |
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clone chad1. |
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Doublets are good! |
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This module parses C<.ace> and related files. A detailed list of methods |
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can be found at the end of this document. |
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I wrote a detailed rationale for design that may explain the reasons |
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why some things were done the way they were done. That document is |
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beyond the scope of this pod and can probably be found in the |
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directory from which this module was 'made' or at |
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L. |
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Note that the POD in that document might be old but the original |
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rationale still stands. |
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=head1 FEEDBACK |
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=head2 Mailing Lists |
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User feedback is an integral part of the evolution of this and other |
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Bioperl modules. Send your comments and suggestions preferably to one |
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of the Bioperl mailing lists. Your participation is much appreciated. |
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bioperl-l@bioperl.org - General discussion |
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http://bioperl.org/wiki/Mailing_lists - About the mailing lists |
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=head2 Support |
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Please direct usage questions or support issues to the mailing list: |
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I |
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rather than to the module maintainer directly. Many experienced and |
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reponsive experts will be able look at the problem and quickly |
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address it. Please include a thorough description of the problem |
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with code and data examples if at all possible. |
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=head2 Reporting Bugs |
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Report bugs to the Bioperl bug tracking system to help us keep track |
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the bugs and their resolution. Bug reports can be submitted via the |
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web: |
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https://github.com/bioperl/bioperl-live/issues |
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=head1 AUTHOR - Chad Matsalla |
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Email chad-at-dieselwurks.com |
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=head1 APPENDIX |
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The rest of the documentation details each of the object |
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methods. Internal methods are usually preceded with a _ |
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=cut |
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#' |
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package Bio::Tools::Alignment::Consed; |
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use strict; |
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use FileHandle; |
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use Dumpvalue qw(dumpValue); |
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use Bio::Tools::Alignment::Trim; |
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use File::Spec; |
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use base qw(Bio::Root::Root Bio::Root::IO); |
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our %DEFAULTS = ( 'f_designator' => 'f', |
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'r_designator' => 'r'); |
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=head2 new() |
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Title : new(-acefile => $path_to_some_acefile, -verbose => "1") |
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Usage : $o_consed = Bio::Tools::Alignment::Consed-> |
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new(-acefile => $path_to_some_acefile, -verbose => "1"); |
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Function: Construct the Bio::Tools::Alignment::Consed object. Sets |
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verbosity for the following procedures, if necessary: |
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1. Construct a new Bio::Tools::Alignment::Trim object, to |
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handle quality trimming 2. Read in the acefile and parse it |
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Returns : A reference to a Bio::Tools::Alignment::Consed object. |
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Args : A hash. (-acefile) is the filename of an acefile. If a full path |
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is not specified "./" is prepended to the filename and used from |
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instantiation until destruction. If you want |
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Bio::Tools::Alignment::Consed to be noisy during parsing of |
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the acefile, specify some value for (-verbose). |
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=cut |
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sub new { |
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my ($class,%args) = @_; |
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my $self = $class->SUPER::new(%args); |
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$self->{'filename'} = $args{'-acefile'}; |
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# this is special to UNIX and should probably use catfile : DONE! |
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# if (!($self->{'filename'} =~ m{/})) { |
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# $self->{'filename'} = "./".$self->{'filename'}; |
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# } |
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# $self->{'filename'} =~ s#\\#\/#g if $^O =~ m/mswin/i; |
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# $self->{'filename'} =~ m/(.*\/)(.*)ace.*$/; |
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# $self->{'path'} = $1; |
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# this is more generic and should work on most systems |
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(undef, $self->{'path'}, undef) = File::Spec->splitpath($self->{'filename'}); |
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$self->_initialize_io('-file'=>$self->{'filename'}); |
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$self->{'o_trim'} = Bio::Tools::Alignment::Trim->new(-verbose => $self->verbose()); |
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$self->set_forward_designator($DEFAULTS{'f_designator'}); |
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$self->set_reverse_designator($DEFAULTS{'r_designator'}); |
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$self->_read_file(); |
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return $self; |
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} |
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168
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=head2 set_verbose() |
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Title : set_verbose() |
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Usage : $o_consed->set_verbose(1); |
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Function: Set the verbosity level for debugging messages. On instantiation |
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of the Bio::Tools::Alignment::Consed object the verbosity level |
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is set to 0 (quiet). |
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Returns : 1 or 0. |
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Args : The verbosity levels are: |
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0 - quiet |
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1 - noisy |
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2 - noisier |
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3 - annoyingly noisy |
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This method for setting verbosity has largely been superseded by a |
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sub-by-sub way, where for every sub you can provide a (-verbose) |
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switch. I am doing converting this bit-by-bit so do not be surprised |
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if some subs do not honour this. |
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187
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=cut |
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189
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# from RootI |
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190
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191
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# backwards compat |
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192
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0
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0
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1
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sub set_verbose { (shift)->verbose(@_) } |
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194
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=head2 get_filename() |
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196
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Title : get_filename() |
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Usage : $o_consed->get_filename(); |
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Function: Returns the name of the acefile being used by the |
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Bio::Tools::Alignment::Consed object. |
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Returns : A scalar containing the name of a file. |
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Args : None. |
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202
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203
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=cut |
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204
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205
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206
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sub get_filename { |
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0
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0
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1
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0
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my $self = shift; |
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0
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0
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return $self->{'filename'}; |
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209
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} |
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210
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211
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=head2 count_sequences_with_grep() |
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212
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213
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Title : count_sequences_with_grep() |
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Usage : $o_consed->count_sequences_with_grep(); |
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215
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Function: Use /bin/grep to scan through the files in the ace project dir |
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216
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and count sequences in those files. I used this method in the |
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217
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development of this module to verify that I was getting all of the |
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218
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sequences. It works, but it is (I think) unix-like platform |
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219
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dependent. |
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Returns : A scalar containing the number of sequences in the ace project |
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221
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directory. |
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Args : None. |
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223
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224
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If you are on a non-UNIX platform, you really do not have to use |
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225
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this. It is more of a debugging routine designed to address very |
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226
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specific problems. |
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227
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228
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This method was reimplemented to be platform independent with a pure |
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229
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perl implementation. The above note can be ignored. |
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230
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231
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=cut |
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232
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233
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sub count_sequences_with_grep { |
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1
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1
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1
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435
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my $self = shift; |
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235
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1
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1
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my ($working_dir,$grep_cli,@total_grep_sequences); |
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236
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# this should be migrated to a pure perl implementation ala |
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237
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# Tom Christiansen's 'tcgrep' |
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238
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# http://www.cpan.org/modules/by-authors/id/TOMC/scripts/tcgrep.gz |
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239
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240
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1
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50
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61
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open my $FILE, '<', $self->{'filename'} or do { |
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241
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0
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0
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$self->warn("Could not read file '$self->{'filename'}' for grepping: $!"); |
|
242
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return |
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243
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0
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0
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}; |
|
244
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1
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2
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my $counter =0; |
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245
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1
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100
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21
|
while(<$FILE>) { $counter++ if(/^AF/); } |
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|
7661
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12680
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246
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1
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10
|
close $FILE; |
|
247
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248
|
1
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46
|
opendir my $SINGLETS, $self->{'path'}; |
|
249
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1
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|
776
|
foreach my $f ( readdir($SINGLETS) ) { |
|
250
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582
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100
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667
|
next unless ($f =~ /\.singlets$/); |
|
251
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252
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1
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20
|
my $singlet_file = File::Spec->catfile($self->{'path'}, $f); |
|
253
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1
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50
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|
32
|
open my $S_FILE, '<', $singlet_file or do { |
|
254
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0
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0
|
$self->warn("Could not read file '$singlet_file': $!"); |
|
255
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|
next |
|
256
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0
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0
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}; |
|
257
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1
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100
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|
24
|
while(<$S_FILE>) { $counter++ if(/^>/) } |
|
|
1291
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2159
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258
|
1
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|
9
|
close $S_FILE; |
|
259
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|
} |
|
260
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1
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|
43
|
closedir $SINGLETS; |
|
261
|
1
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|
8
|
return $counter; |
|
262
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|
} |
|
263
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|
264
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|
=head2 get_path() |
|
265
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|
266
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|
|
Title : get_path() |
|
267
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|
|
Usage : $o_consed->get_path(); |
|
268
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|
Function: Returns the path to the acefile this object is working with. |
|
269
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|
|
Returns : Scalar. The path to the working acefile. |
|
270
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|
Args : None. |
|
271
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|
272
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|
=cut |
|
273
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|
274
|
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|
|
sub get_path { |
|
275
|
0
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|
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0
|
1
|
0
|
my $self = shift; |
|
276
|
0
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|
|
|
|
0
|
return $self->{'path'}; |
|
277
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|
|
} |
|
278
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|
279
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|
|
=head2 get_contigs() |
|
280
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|
281
|
|
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|
|
Title : get_contigs() |
|
282
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|
|
Usage : $o_consed->get_contigs(); |
|
283
|
|
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|
|
Function: Return the keys to the Bio::Tools::Alignment::Consed object. |
|
284
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|
|
|
|
|
|
Returns : An array containing the keynames in the |
|
285
|
|
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|
|
|
|
Bio::Tools::Alignment::Consed object. |
|
286
|
|
|
|
|
|
|
Args : None. |
|
287
|
|
|
|
|
|
|
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|
288
|
|
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|
|
This would normally be used to get the keynames for some sort of |
|
289
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|
|
|
|
|
|
iterator. These keys are worthless in general day-to-day use because |
|
290
|
|
|
|
|
|
|
in the Consed acefile they are simply Contig1, Contig2, ... |
|
291
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|
292
|
|
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|
|
=cut |
|
293
|
|
|
|
|
|
|
|
|
294
|
|
|
|
|
|
|
sub get_contigs { |
|
295
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
296
|
0
|
|
|
|
|
0
|
my @contigs = sort keys %{$self->{'contigs'}}; |
|
|
0
|
|
|
|
|
0
|
|
|
297
|
0
|
|
|
|
|
0
|
return @contigs; |
|
298
|
|
|
|
|
|
|
} |
|
299
|
|
|
|
|
|
|
|
|
300
|
|
|
|
|
|
|
=head2 get_class($contig_keyname) |
|
301
|
|
|
|
|
|
|
|
|
302
|
|
|
|
|
|
|
Title : get_class($contig_keyname) |
|
303
|
|
|
|
|
|
|
Usage : $o_consed->get_class($contig_keyname); |
|
304
|
|
|
|
|
|
|
Function: Return the class name for this contig |
|
305
|
|
|
|
|
|
|
Returns : A scalar representing the class of this contig. |
|
306
|
|
|
|
|
|
|
Args : None. |
|
307
|
|
|
|
|
|
|
Notes : |
|
308
|
|
|
|
|
|
|
|
|
309
|
|
|
|
|
|
|
=cut |
|
310
|
|
|
|
|
|
|
|
|
311
|
|
|
|
|
|
|
sub get_class { |
|
312
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
313
|
0
|
|
|
|
|
0
|
return $self->{contigs}->{$contig}->{class}; |
|
314
|
|
|
|
|
|
|
} |
|
315
|
|
|
|
|
|
|
|
|
316
|
|
|
|
|
|
|
=head2 get_quality_array($contig_keyname) |
|
317
|
|
|
|
|
|
|
|
|
318
|
|
|
|
|
|
|
Title : get_quality_array($contig_keyname) |
|
319
|
|
|
|
|
|
|
Usage : $o_consed->get_quality_array($contig_keyname); |
|
320
|
|
|
|
|
|
|
Function: Returns the quality for the consensus sequence for the given |
|
321
|
|
|
|
|
|
|
contig as an array. See get_quality_scalar to get this as a scalar. |
|
322
|
|
|
|
|
|
|
Returns : An array containing the quality for the consensus sequence with |
|
323
|
|
|
|
|
|
|
the given keyname. |
|
324
|
|
|
|
|
|
|
Args : The keyname of a contig. Note: This is a keyname. The key would |
|
325
|
|
|
|
|
|
|
normally come from get_contigs. |
|
326
|
|
|
|
|
|
|
|
|
327
|
|
|
|
|
|
|
Returns an array, not a reference. Is this a bug? I No. |
|
328
|
|
|
|
|
|
|
Well, maybe. Why was this developed like this? I was using FreezeThaw |
|
329
|
|
|
|
|
|
|
for object persistence, and when it froze out these arrays it took a |
|
330
|
|
|
|
|
|
|
long time to thaw it. Much better as a scalar. |
|
331
|
|
|
|
|
|
|
|
|
332
|
|
|
|
|
|
|
See L |
|
333
|
|
|
|
|
|
|
|
|
334
|
|
|
|
|
|
|
=cut |
|
335
|
|
|
|
|
|
|
|
|
336
|
|
|
|
|
|
|
sub get_quality_array { |
|
337
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
338
|
0
|
|
|
|
|
0
|
return split ' ', $self->{contigs}->{$contig}->{quality}; |
|
339
|
|
|
|
|
|
|
} |
|
340
|
|
|
|
|
|
|
|
|
341
|
|
|
|
|
|
|
=head2 get_quality_scalar($contig_keyname) |
|
342
|
|
|
|
|
|
|
|
|
343
|
|
|
|
|
|
|
Title : get_quality_scalar($contig_keyname) |
|
344
|
|
|
|
|
|
|
Usage : $o_consed->get_quality_scalar($contig_keyname); |
|
345
|
|
|
|
|
|
|
Function: Returns the quality for the consensus sequence for the given |
|
346
|
|
|
|
|
|
|
contig as a scalar. See get_quality_array to get this as an array. |
|
347
|
|
|
|
|
|
|
Returns : An scalar containing the quality for the consensus sequence with |
|
348
|
|
|
|
|
|
|
the given keyname. |
|
349
|
|
|
|
|
|
|
Args : The keyname of a contig. Note this is a _keyname_. The key would |
|
350
|
|
|
|
|
|
|
normally come from get_contigs. |
|
351
|
|
|
|
|
|
|
|
|
352
|
|
|
|
|
|
|
Why was this developed like this? I was using FreezeThaw for object |
|
353
|
|
|
|
|
|
|
persistence, and when it froze out these arrays it took a coon's age |
|
354
|
|
|
|
|
|
|
to thaw it. Much better as a scalar. |
|
355
|
|
|
|
|
|
|
|
|
356
|
|
|
|
|
|
|
See L |
|
357
|
|
|
|
|
|
|
|
|
358
|
|
|
|
|
|
|
=cut |
|
359
|
|
|
|
|
|
|
|
|
360
|
|
|
|
|
|
|
#' |
|
361
|
|
|
|
|
|
|
sub get_quality_scalar { |
|
362
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
363
|
0
|
|
|
|
|
0
|
return $self->{'contigs'}->{$contig}->{'quality'}; |
|
364
|
|
|
|
|
|
|
} |
|
365
|
|
|
|
|
|
|
|
|
366
|
|
|
|
|
|
|
=head2 freeze_hash() |
|
367
|
|
|
|
|
|
|
|
|
368
|
|
|
|
|
|
|
Title : freeze_hash() |
|
369
|
|
|
|
|
|
|
Usage : $o_consed->freeze_hash(); |
|
370
|
|
|
|
|
|
|
|
|
371
|
|
|
|
|
|
|
Function: Use Ilya's FreezeThaw module to create a persistent data |
|
372
|
|
|
|
|
|
|
object for this Bio::Tools::Alignment::Consed data |
|
373
|
|
|
|
|
|
|
structure. In the case of AAFC, we use |
|
374
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed to pre-process bunches of |
|
375
|
|
|
|
|
|
|
sequences, freeze the structures, and send in a harvesting |
|
376
|
|
|
|
|
|
|
robot later to do database stuff. |
|
377
|
|
|
|
|
|
|
Returns : 0 or 1; |
|
378
|
|
|
|
|
|
|
Args : None. |
|
379
|
|
|
|
|
|
|
|
|
380
|
|
|
|
|
|
|
This procedure was removed so Consed.pm won't require FreezeThaw. |
|
381
|
|
|
|
|
|
|
|
|
382
|
|
|
|
|
|
|
=cut |
|
383
|
|
|
|
|
|
|
|
|
384
|
|
|
|
|
|
|
#' |
|
385
|
|
|
|
|
|
|
sub freeze_hash { |
|
386
|
0
|
|
|
0
|
1
|
0
|
my $self = shift; |
|
387
|
0
|
|
|
|
|
0
|
$self->warn("This method (freeze_hash) was removed ". |
|
388
|
|
|
|
|
|
|
"from the bioperl consed.pm. Sorry.\n"); |
|
389
|
0
|
|
|
|
|
0
|
if (1==2) { |
|
390
|
|
|
|
|
|
|
$self->debug("Bio::Tools::Alignment::Consed::freeze_hash:". |
|
391
|
|
|
|
|
|
|
" \$self->{path} is $self->{path}\n"); |
|
392
|
|
|
|
|
|
|
my $filename = $self->{'path'}."frozen"; |
|
393
|
|
|
|
|
|
|
my %contigs = %{$self->{'contigs'}}; |
|
394
|
|
|
|
|
|
|
my $frozen = freeze(%contigs); |
|
395
|
|
|
|
|
|
|
umask 0001; |
|
396
|
|
|
|
|
|
|
open my $FREEZE, '>', $filename or do { |
|
397
|
|
|
|
|
|
|
$self->warn( "Bio::Tools::Alignment::Consed could not ". |
|
398
|
|
|
|
|
|
|
"freeze the contig hash because the file ". |
|
399
|
|
|
|
|
|
|
"($filename) could not be opened: $!"); |
|
400
|
|
|
|
|
|
|
return 1; |
|
401
|
|
|
|
|
|
|
}; |
|
402
|
|
|
|
|
|
|
print $FREEZE $frozen; |
|
403
|
|
|
|
|
|
|
return 0; |
|
404
|
|
|
|
|
|
|
} |
|
405
|
|
|
|
|
|
|
} |
|
406
|
|
|
|
|
|
|
|
|
407
|
|
|
|
|
|
|
=head2 get_members($contig_keyname) |
|
408
|
|
|
|
|
|
|
|
|
409
|
|
|
|
|
|
|
Title : get_members($contig_keyname) |
|
410
|
|
|
|
|
|
|
Usage : $o_consed->get_members($contig_keyname); |
|
411
|
|
|
|
|
|
|
Function: Return the _names_ of the reads in this contig. |
|
412
|
|
|
|
|
|
|
Returns : An array containing the names of the reads in this contig. |
|
413
|
|
|
|
|
|
|
Args : The keyname of a contig. Note this is a keyname. The keyname |
|
414
|
|
|
|
|
|
|
would normally come from get_contigs. |
|
415
|
|
|
|
|
|
|
|
|
416
|
|
|
|
|
|
|
See L |
|
417
|
|
|
|
|
|
|
|
|
418
|
|
|
|
|
|
|
=cut |
|
419
|
|
|
|
|
|
|
|
|
420
|
|
|
|
|
|
|
sub get_members { |
|
421
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
422
|
0
|
0
|
|
|
|
0
|
if (!$contig) { |
|
423
|
0
|
|
|
|
|
0
|
$self->warn("You need to provide the name of a contig to ". |
|
424
|
|
|
|
|
|
|
"use Bio::Tools::Alignment::Consed::get_members!\n"); |
|
425
|
0
|
|
|
|
|
0
|
return; |
|
426
|
|
|
|
|
|
|
} |
|
427
|
0
|
|
|
|
|
0
|
return @{$self->{'contigs'}->{$contig}->{'member_array'}}; |
|
|
0
|
|
|
|
|
0
|
|
|
428
|
|
|
|
|
|
|
} |
|
429
|
|
|
|
|
|
|
|
|
430
|
|
|
|
|
|
|
=head2 get_members_by_name($some_arbitrary_name) |
|
431
|
|
|
|
|
|
|
|
|
432
|
|
|
|
|
|
|
Title : get_members_by_name($some_arbitrary_name) |
|
433
|
|
|
|
|
|
|
Usage : $o_consed->get_members_by_name($some_arbitrary_name); |
|
434
|
|
|
|
|
|
|
Function: Return the names of the reads in a contig. This is the name given |
|
435
|
|
|
|
|
|
|
to $contig{key} based on what is in the contig. This is different |
|
436
|
|
|
|
|
|
|
from the keys retrieved through get_contigs(). |
|
437
|
|
|
|
|
|
|
Returns : An array containing the names of the reads in the contig with this |
|
438
|
|
|
|
|
|
|
name. |
|
439
|
|
|
|
|
|
|
Args : The name of a contig. Not a key, but a name. |
|
440
|
|
|
|
|
|
|
|
|
441
|
|
|
|
|
|
|
Highly inefficient. use some other method if possible. |
|
442
|
|
|
|
|
|
|
See L |
|
443
|
|
|
|
|
|
|
|
|
444
|
|
|
|
|
|
|
=cut |
|
445
|
|
|
|
|
|
|
|
|
446
|
|
|
|
|
|
|
sub get_members_by_name { |
|
447
|
0
|
|
|
0
|
1
|
0
|
my ($self,$name) = @_; |
|
448
|
|
|
|
|
|
|
# build a list to try to screen for redundancy |
|
449
|
0
|
|
|
|
|
0
|
my @contigs_with_that_name; |
|
450
|
0
|
|
|
|
|
0
|
foreach my $currkey ( sort keys %{$self->{'contigs'}} ) { |
|
|
0
|
|
|
|
|
0
|
|
|
451
|
0
|
0
|
|
|
|
0
|
next if (!$self->{'contigs'}->{$currkey}->{'name'}); |
|
452
|
0
|
0
|
|
|
|
0
|
if ($self->{'contigs'}->{$currkey}->{'name'} eq "$name") { |
|
453
|
0
|
|
|
|
|
0
|
push @contigs_with_that_name,$currkey; |
|
454
|
|
|
|
|
|
|
} |
|
455
|
|
|
|
|
|
|
} |
|
456
|
0
|
|
|
|
|
0
|
my $count = @contigs_with_that_name; |
|
457
|
0
|
0
|
|
|
|
0
|
if ($count == 1) { |
|
458
|
0
|
|
|
|
|
0
|
my $contig_num = $contigs_with_that_name[0]; |
|
459
|
0
|
|
|
|
|
0
|
return @{$self->{'contigs'}->{$contig_num}->{'member_array'}}; |
|
|
0
|
|
|
|
|
0
|
|
|
460
|
|
|
|
|
|
|
} |
|
461
|
|
|
|
|
|
|
} |
|
462
|
|
|
|
|
|
|
|
|
463
|
|
|
|
|
|
|
=head2 get_contig_number_by_name($some_arbitrary_name) |
|
464
|
|
|
|
|
|
|
|
|
465
|
|
|
|
|
|
|
Title : get_contig_number_by_name($some_arbitrary_name) |
|
466
|
|
|
|
|
|
|
Usage : $o_consed->get_contig_number_by_name($some_arbitrary_name); |
|
467
|
|
|
|
|
|
|
Function: Return the names of the reads in a contig. This is the name given |
|
468
|
|
|
|
|
|
|
to $contig{key} based on what is in the contig. This is different |
|
469
|
|
|
|
|
|
|
from the keys retrieved through get_contigs(). |
|
470
|
|
|
|
|
|
|
Returns : An array containing the names of the reads in the contig with this |
|
471
|
|
|
|
|
|
|
name. |
|
472
|
|
|
|
|
|
|
Args : The name of a contig. Not a key, but a name. |
|
473
|
|
|
|
|
|
|
|
|
474
|
|
|
|
|
|
|
See L |
|
475
|
|
|
|
|
|
|
|
|
476
|
|
|
|
|
|
|
=cut |
|
477
|
|
|
|
|
|
|
|
|
478
|
|
|
|
|
|
|
sub get_contig_number_by_name { |
|
479
|
0
|
|
|
0
|
1
|
0
|
my ($self,$name) = @_; |
|
480
|
0
|
|
|
|
|
0
|
foreach my $currkey (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
0
|
|
|
481
|
0
|
0
|
0
|
|
|
0
|
if ($self->{'contigs'}->{$currkey}->{'name'} && |
|
482
|
|
|
|
|
|
|
$self->{'contigs'}->{$currkey}->{'name'} eq "$name") { |
|
483
|
0
|
|
|
|
|
0
|
return $currkey; |
|
484
|
|
|
|
|
|
|
} |
|
485
|
|
|
|
|
|
|
} |
|
486
|
|
|
|
|
|
|
} |
|
487
|
|
|
|
|
|
|
|
|
488
|
|
|
|
|
|
|
=head2 get_sequence($contig_keyname) |
|
489
|
|
|
|
|
|
|
|
|
490
|
|
|
|
|
|
|
Title : get_sequence($contig_keyname) |
|
491
|
|
|
|
|
|
|
Usage : $o_consed->get_sequence($contig_keyname); |
|
492
|
|
|
|
|
|
|
Function: Returns the consensus sequence for a given contig. |
|
493
|
|
|
|
|
|
|
Returns : A scalar containing a sequence. |
|
494
|
|
|
|
|
|
|
Args : The keyname of a contig. Note this is a key. The key would |
|
495
|
|
|
|
|
|
|
normally come from get_contigs. |
|
496
|
|
|
|
|
|
|
|
|
497
|
|
|
|
|
|
|
See L |
|
498
|
|
|
|
|
|
|
|
|
499
|
|
|
|
|
|
|
=cut |
|
500
|
|
|
|
|
|
|
|
|
501
|
|
|
|
|
|
|
sub get_sequence { |
|
502
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
503
|
0
|
|
|
|
|
0
|
return $self->{'contigs'}->{$contig}->{'consensus'}; |
|
504
|
|
|
|
|
|
|
} |
|
505
|
|
|
|
|
|
|
|
|
506
|
|
|
|
|
|
|
=head2 set_final_sequence($some_sequence) |
|
507
|
|
|
|
|
|
|
|
|
508
|
|
|
|
|
|
|
Title : set_final_sequence($name,$some_sequence) |
|
509
|
|
|
|
|
|
|
Usage : $o_consed->set_final_sequence($name,$some_sequence); |
|
510
|
|
|
|
|
|
|
Function: Provides a manual way to set the sequence for a given key in the |
|
511
|
|
|
|
|
|
|
contig hash. Rarely used. |
|
512
|
|
|
|
|
|
|
Returns : 0 or 1; |
|
513
|
|
|
|
|
|
|
Args : The name (not the keyname) of a contig and an arbitrary string. |
|
514
|
|
|
|
|
|
|
|
|
515
|
|
|
|
|
|
|
A method with a questionable and somewhat mysterious origin. May raise |
|
516
|
|
|
|
|
|
|
the dead or something like that. |
|
517
|
|
|
|
|
|
|
|
|
518
|
|
|
|
|
|
|
=cut |
|
519
|
|
|
|
|
|
|
|
|
520
|
|
|
|
|
|
|
sub set_final_sequence { |
|
521
|
0
|
|
|
0
|
1
|
0
|
my ($self,$name,$sequence) = @_; |
|
522
|
0
|
0
|
|
|
|
0
|
if (!$self->{'contigs'}->{$name}) { |
|
523
|
0
|
|
|
|
|
0
|
$self->warn("You cannot set the final sequence for ". |
|
524
|
|
|
|
|
|
|
"$name because it doesn't exist!\n"); |
|
525
|
0
|
|
|
|
|
0
|
return 1; |
|
526
|
|
|
|
|
|
|
} |
|
527
|
|
|
|
|
|
|
else { |
|
528
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$name}->{'final_sequence'} = $sequence; |
|
529
|
|
|
|
|
|
|
} |
|
530
|
0
|
|
|
|
|
0
|
return 0; |
|
531
|
|
|
|
|
|
|
} |
|
532
|
|
|
|
|
|
|
|
|
533
|
|
|
|
|
|
|
=head2 _read_file() |
|
534
|
|
|
|
|
|
|
|
|
535
|
|
|
|
|
|
|
Title : _read_file(); |
|
536
|
|
|
|
|
|
|
Usage : _read_file(); |
|
537
|
|
|
|
|
|
|
Function: An internal subroutine used to read in an acefile and parse it |
|
538
|
|
|
|
|
|
|
into a Bio::Tools::Alignment::Consed object. |
|
539
|
|
|
|
|
|
|
Returns : 0 or 1. |
|
540
|
|
|
|
|
|
|
Args : Nothing. |
|
541
|
|
|
|
|
|
|
|
|
542
|
|
|
|
|
|
|
This routine creates and saves the filhandle for reading the files in |
|
543
|
|
|
|
|
|
|
{fh} |
|
544
|
|
|
|
|
|
|
|
|
545
|
|
|
|
|
|
|
=cut |
|
546
|
|
|
|
|
|
|
|
|
547
|
|
|
|
|
|
|
sub _read_file { |
|
548
|
1
|
|
|
1
|
|
1
|
my ($self) = @_; |
|
549
|
1
|
|
|
|
|
1
|
my ($line,$in_contig,$in_quality,$contig_number,$top); |
|
550
|
|
|
|
|
|
|
# make it easier to type $fhl |
|
551
|
1
|
|
|
|
|
5
|
while (defined($line=$self->_readline()) ) { |
|
552
|
7661
|
|
|
|
|
5578
|
chomp $line; |
|
553
|
|
|
|
|
|
|
# check if there is anything on this line |
|
554
|
|
|
|
|
|
|
# if not, you can stop gathering consensus sequence |
|
555
|
7661
|
100
|
|
|
|
23701
|
if (!$line) { |
|
|
|
100
|
|
|
|
|
|
|
|
|
100
|
|
|
|
|
|
|
|
|
100
|
|
|
|
|
|
|
|
|
100
|
|
|
|
|
|
|
|
|
50
|
|
|
|
|
|
|
|
|
100
|
|
|
|
|
|
|
556
|
|
|
|
|
|
|
# if the line is blank you are no longer to gather consensus |
|
557
|
|
|
|
|
|
|
# sequence or quality values |
|
558
|
488
|
|
|
|
|
371
|
$in_contig = 0; |
|
559
|
488
|
|
|
|
|
627
|
$in_quality = 0; |
|
560
|
|
|
|
|
|
|
} |
|
561
|
|
|
|
|
|
|
# you are currently gathering consensus sequence |
|
562
|
|
|
|
|
|
|
elsif ($in_contig) { |
|
563
|
910
|
50
|
|
|
|
1033
|
if ($in_contig == 1) { |
|
564
|
910
|
|
|
|
|
1775
|
$self->debug("Adding $line to consensus of contig number $contig_number.\n"); |
|
565
|
910
|
|
|
|
|
1971
|
$self->{'contigs'}->{$contig_number}->{'consensus'} .= $line; |
|
566
|
|
|
|
|
|
|
} |
|
567
|
|
|
|
|
|
|
} |
|
568
|
|
|
|
|
|
|
elsif ($in_quality) { |
|
569
|
910
|
50
|
|
|
|
767
|
if (!$line) { |
|
570
|
0
|
|
|
|
|
0
|
$in_quality = undef; |
|
571
|
|
|
|
|
|
|
} |
|
572
|
|
|
|
|
|
|
else { |
|
573
|
|
|
|
|
|
|
|
|
574
|
|
|
|
|
|
|
# I wrote this in here because acefiles produced by |
|
575
|
|
|
|
|
|
|
# cap3 do not have a leading space like the acefiles |
|
576
|
|
|
|
|
|
|
# produced by phrap and there is the potential to have |
|
577
|
|
|
|
|
|
|
# concatenated quality values like this: 2020 rather |
|
578
|
|
|
|
|
|
|
# then 20 20 whre lines collide. Thanks Andrew for |
|
579
|
|
|
|
|
|
|
# noticing. |
|
580
|
|
|
|
|
|
|
|
|
581
|
910
|
100
|
66
|
|
|
2608
|
if ($self->{'contigs'}->{$contig_number}->{'quality'} && |
|
582
|
|
|
|
|
|
|
!($self->{'contigs'}->{$contig_number}->{'quality'} =~ m/\ $/)) { |
|
583
|
857
|
|
|
|
|
738
|
$self->{'contigs'}->{$contig_number}->{'quality'} .= " "; |
|
584
|
|
|
|
|
|
|
} |
|
585
|
910
|
|
|
|
|
1843
|
$self->{'contigs'}->{$contig_number}->{'quality'} .= $line; |
|
586
|
|
|
|
|
|
|
} |
|
587
|
|
|
|
|
|
|
} |
|
588
|
|
|
|
|
|
|
elsif ($line =~ /^BQ/) { |
|
589
|
53
|
|
|
|
|
84
|
$in_quality = 1; |
|
590
|
|
|
|
|
|
|
} |
|
591
|
|
|
|
|
|
|
|
|
592
|
|
|
|
|
|
|
# the line /^CO/ like this: |
|
593
|
|
|
|
|
|
|
# CO Contig1 796 1 1 U |
|
594
|
|
|
|
|
|
|
# can be broken down as follows: |
|
595
|
|
|
|
|
|
|
# CO - Contig! |
|
596
|
|
|
|
|
|
|
# Contig1 - the name of this contig |
|
597
|
|
|
|
|
|
|
# 796 - Number of bases in this contig |
|
598
|
|
|
|
|
|
|
# 1 - Number of reads in this contig |
|
599
|
|
|
|
|
|
|
# 1 - number of base segments in this contig |
|
600
|
|
|
|
|
|
|
# U - Uncomplemented |
|
601
|
|
|
|
|
|
|
|
|
602
|
|
|
|
|
|
|
elsif ($line =~ /^CO/) { |
|
603
|
53
|
|
|
|
|
114
|
$line =~ m/^CO\ Contig(\d+)\ \d+\ \d+\ \d+\ (\w)/; |
|
604
|
53
|
|
|
|
|
78
|
$contig_number = $1; |
|
605
|
53
|
50
|
|
|
|
93
|
if ($2 eq "C") { |
|
606
|
0
|
|
|
|
|
0
|
$self->debug("Contig $contig_number is complemented!\n"); |
|
607
|
|
|
|
|
|
|
} |
|
608
|
53
|
|
|
|
|
144
|
$self->{'contigs'}->{$contig_number}->{'member_array'} = []; |
|
609
|
53
|
|
|
|
|
89
|
$self->{'contigs'}->{$contig_number}->{'contig_direction'} = "$2"; |
|
610
|
53
|
|
|
|
|
97
|
$in_contig = 1; |
|
611
|
|
|
|
|
|
|
} |
|
612
|
|
|
|
|
|
|
|
|
613
|
|
|
|
|
|
|
# 000713 |
|
614
|
|
|
|
|
|
|
# this BS is deprecated, I think. |
|
615
|
|
|
|
|
|
|
# haha, I am really witty. |
|
616
|
|
|
|
|
|
|
|
|
617
|
|
|
|
|
|
|
elsif ($line =~ /^BSDEPRECATED/) { |
|
618
|
0
|
|
|
|
|
0
|
$line =~ m/^BS\s+\d+\s+\d+\s+(.+)/; |
|
619
|
0
|
|
|
|
|
0
|
my $member = $1; |
|
620
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{$member}++; |
|
621
|
|
|
|
|
|
|
} |
|
622
|
|
|
|
|
|
|
# the members of the contigs are determined by the AF line in the ace file |
|
623
|
|
|
|
|
|
|
elsif ($line =~ /^AF/) { |
|
624
|
114
|
|
|
|
|
209
|
$self->debug("I see an AF line here.\n"); |
|
625
|
114
|
|
|
|
|
255
|
$line =~ /^AF\ (\S+)\ (\w)\ (\S+)/; |
|
626
|
|
|
|
|
|
|
|
|
627
|
|
|
|
|
|
|
# push the name of the current read onto the member array for this contig |
|
628
|
114
|
|
|
|
|
77
|
push @{$self->{'contigs'}->{$contig_number}->{'member_array'}},$1; |
|
|
114
|
|
|
|
|
287
|
|
|
629
|
|
|
|
|
|
|
|
|
630
|
|
|
|
|
|
|
# the first read in the contig will be named the "top" read |
|
631
|
114
|
100
|
|
|
|
137
|
if (!$top) { |
|
632
|
57
|
|
|
|
|
77
|
$self->debug("\$top is not set.\n"); |
|
633
|
57
|
50
|
|
|
|
82
|
if ($self->{'contigs'}->{$contig_number}->{'contig_direction'} eq "C") { |
|
634
|
0
|
|
|
|
|
0
|
$self->debug("Reversing the order of the reads. The bottom will be $1\n"); |
|
635
|
|
|
|
|
|
|
|
|
636
|
|
|
|
|
|
|
# if the contig sequence is marked as the |
|
637
|
|
|
|
|
|
|
# complement, the top becomes the bottom and$ |
|
638
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'bottom_name'} = $1; |
|
639
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'bottom_complement'} = $2; |
|
640
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'bottom_start'} = $3; |
|
641
|
|
|
|
|
|
|
} |
|
642
|
|
|
|
|
|
|
else { |
|
643
|
57
|
|
|
|
|
148
|
$self->debug("NOT reversing the order of the reads. ". |
|
644
|
|
|
|
|
|
|
"The top_name will be $1\n"); |
|
645
|
|
|
|
|
|
|
# if the contig sequence is marked as the |
|
646
|
|
|
|
|
|
|
# complement, the top becomes the bottom and$ |
|
647
|
57
|
|
|
|
|
140
|
$self->{'contigs'}->{$contig_number}->{'top_name'} = $1; |
|
648
|
57
|
|
|
|
|
75
|
$self->{'contigs'}->{$contig_number}->{'top_complement'} = $2; |
|
649
|
57
|
|
|
|
|
79
|
$self->{'contigs'}->{$contig_number}->{'top_start'} = $3; |
|
650
|
|
|
|
|
|
|
} |
|
651
|
57
|
|
|
|
|
93
|
$top = 1; |
|
652
|
|
|
|
|
|
|
} |
|
653
|
|
|
|
|
|
|
else { |
|
654
|
|
|
|
|
|
|
|
|
655
|
|
|
|
|
|
|
# if the contig sequence is marked as the complement, |
|
656
|
|
|
|
|
|
|
# the top becomes the bottom and the bottom becomes |
|
657
|
|
|
|
|
|
|
# the top |
|
658
|
57
|
50
|
|
|
|
83
|
if ($self->{'contigs'}->{$contig_number}->{'contig_direction'} eq "C") { |
|
659
|
0
|
|
|
|
|
0
|
$self->debug("Reversing the order of the reads. The top will be $1\n"); |
|
660
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'top_name'} = $1; |
|
661
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'top_complement'} = $2; |
|
662
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$contig_number}->{'top_start'} = $3; |
|
663
|
|
|
|
|
|
|
} |
|
664
|
|
|
|
|
|
|
else { |
|
665
|
57
|
|
|
|
|
135
|
$self->debug("NOT reversing the order of the reads. The bottom will be $1\n"); |
|
666
|
57
|
|
|
|
|
100
|
$self->{'contigs'}->{$contig_number}->{'bottom_name'} = $1; |
|
667
|
57
|
|
|
|
|
89
|
$self->{'contigs'}->{$contig_number}->{'bottom_complement'} = $2; |
|
668
|
57
|
|
|
|
|
92
|
$self->{'contigs'}->{$contig_number}->{'bottom_start'} = $3; |
|
669
|
|
|
|
|
|
|
} |
|
670
|
57
|
|
|
|
|
101
|
$top = undef; |
|
671
|
|
|
|
|
|
|
} |
|
672
|
|
|
|
|
|
|
} |
|
673
|
|
|
|
|
|
|
} |
|
674
|
1
|
|
|
|
|
2
|
return 0; |
|
675
|
|
|
|
|
|
|
} |
|
676
|
|
|
|
|
|
|
|
|
677
|
|
|
|
|
|
|
=head2 set_reverse_designator($some_string) |
|
678
|
|
|
|
|
|
|
|
|
679
|
|
|
|
|
|
|
Title : set_reverse_designator($some_string) |
|
680
|
|
|
|
|
|
|
Usage : $o_consed->set_reverse_designator($some_string); |
|
681
|
|
|
|
|
|
|
Function: Set the designator for the reverse read of contigs in this |
|
682
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. Used to determine if |
|
683
|
|
|
|
|
|
|
contigs containing two reads can be named. |
|
684
|
|
|
|
|
|
|
Returns : The value of $o_consed->{reverse_designator} so you can check |
|
685
|
|
|
|
|
|
|
to see that it was set properly. |
|
686
|
|
|
|
|
|
|
Args : An arbitrary string. |
|
687
|
|
|
|
|
|
|
|
|
688
|
|
|
|
|
|
|
May be useful only to me. I |
|
689
|
|
|
|
|
|
|
|
|
690
|
|
|
|
|
|
|
=cut |
|
691
|
|
|
|
|
|
|
|
|
692
|
|
|
|
|
|
|
sub set_reverse_designator { |
|
693
|
1
|
|
|
1
|
1
|
1
|
my ($self,$reverse_designator) = @_; |
|
694
|
1
|
|
|
|
|
2
|
$self->{'reverse_designator'} = $reverse_designator; |
|
695
|
1
|
|
|
|
|
3
|
$self->{'o_trim'}->set_reverse_designator($reverse_designator); |
|
696
|
1
|
|
|
|
|
1
|
return $self->{'reverse_designator'}; |
|
697
|
|
|
|
|
|
|
} # end set_reverse_designator |
|
698
|
|
|
|
|
|
|
|
|
699
|
|
|
|
|
|
|
=head2 set_forward_designator($some_string) |
|
700
|
|
|
|
|
|
|
|
|
701
|
|
|
|
|
|
|
Title : set_forward_designator($some_string) |
|
702
|
|
|
|
|
|
|
Usage : $o_consed->set_forward_designator($some_string); |
|
703
|
|
|
|
|
|
|
Function: Set the designator for the forward read of contigs in this |
|
704
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. Used to determine if |
|
705
|
|
|
|
|
|
|
contigs containing two reads can be named. |
|
706
|
|
|
|
|
|
|
Returns : The value of $o_consed->{forward_designator} so you can check |
|
707
|
|
|
|
|
|
|
to see that it was set properly. |
|
708
|
|
|
|
|
|
|
Args : An arbitrary string. |
|
709
|
|
|
|
|
|
|
|
|
710
|
|
|
|
|
|
|
May be useful only to me. I |
|
711
|
|
|
|
|
|
|
|
|
712
|
|
|
|
|
|
|
=cut |
|
713
|
|
|
|
|
|
|
|
|
714
|
|
|
|
|
|
|
sub set_forward_designator { |
|
715
|
1
|
|
|
1
|
1
|
2
|
my ($self,$forward_designator) = @_; |
|
716
|
1
|
|
|
|
|
2
|
$self->{'forward_designator'} = $forward_designator; |
|
717
|
1
|
|
|
|
|
2
|
$self->{'o_trim'}->set_forward_designator($forward_designator); |
|
718
|
1
|
|
|
|
|
1
|
return $self->{'forward_designator'}; |
|
719
|
|
|
|
|
|
|
} # end set_forward_designator |
|
720
|
|
|
|
|
|
|
|
|
721
|
|
|
|
|
|
|
=head2 set_designator_ignore_case("yes") |
|
722
|
|
|
|
|
|
|
|
|
723
|
|
|
|
|
|
|
Title : set_designator_ignore_case("yes") |
|
724
|
|
|
|
|
|
|
Usage : $o_consed->set_designator_ignore_case("yes"); |
|
725
|
|
|
|
|
|
|
Function: Deprecated. |
|
726
|
|
|
|
|
|
|
Returns : Deprecated. |
|
727
|
|
|
|
|
|
|
Args : Deprecated. |
|
728
|
|
|
|
|
|
|
|
|
729
|
|
|
|
|
|
|
Deprecated. Really. Trust me. |
|
730
|
|
|
|
|
|
|
|
|
731
|
|
|
|
|
|
|
=cut |
|
732
|
|
|
|
|
|
|
|
|
733
|
|
|
|
|
|
|
sub set_designator_ignore_case { |
|
734
|
0
|
|
|
0
|
1
|
0
|
my ($self,$ignore_case) = @_; |
|
735
|
0
|
0
|
|
|
|
0
|
if ($ignore_case eq "yes") { |
|
736
|
0
|
|
|
|
|
0
|
$self->{'designator_ignore_case'} = 1; |
|
737
|
|
|
|
|
|
|
} |
|
738
|
0
|
|
|
|
|
0
|
return $self->{'designator_ignore_case'}; |
|
739
|
|
|
|
|
|
|
} # end set_designator_ignore_case |
|
740
|
|
|
|
|
|
|
|
|
741
|
|
|
|
|
|
|
=head2 set_trim_points_singlets_and_singletons() |
|
742
|
|
|
|
|
|
|
|
|
743
|
|
|
|
|
|
|
Title : set_trim_points_singlets_and_singletons() |
|
744
|
|
|
|
|
|
|
Usage : $o_consed->set_trim_points_singlets_and_singletons(); |
|
745
|
|
|
|
|
|
|
Function: Set the trim points for singlets and singletons based on |
|
746
|
|
|
|
|
|
|
quality. Uses the Bio::Tools::Alignment::Trim object. Use |
|
747
|
|
|
|
|
|
|
at your own risk because the Bio::Tools::Alignment::Trim |
|
748
|
|
|
|
|
|
|
object was designed specifically for me and is mysterious |
|
749
|
|
|
|
|
|
|
in its ways. Every time somebody other then me uses it a |
|
750
|
|
|
|
|
|
|
swarm of locusts decends on a small Central American |
|
751
|
|
|
|
|
|
|
village so do not say you weren't warned. |
|
752
|
|
|
|
|
|
|
Returns : Nothing. |
|
753
|
|
|
|
|
|
|
Args : None. |
|
754
|
|
|
|
|
|
|
|
|
755
|
|
|
|
|
|
|
Working on exceptions and warnings here. |
|
756
|
|
|
|
|
|
|
|
|
757
|
|
|
|
|
|
|
See L for more information |
|
758
|
|
|
|
|
|
|
|
|
759
|
|
|
|
|
|
|
=cut |
|
760
|
|
|
|
|
|
|
|
|
761
|
|
|
|
|
|
|
#' to make my emacs happy |
|
762
|
|
|
|
|
|
|
|
|
763
|
|
|
|
|
|
|
sub set_trim_points_singlets_and_singletons { |
|
764
|
0
|
|
|
0
|
1
|
0
|
my ($self) = @_; |
|
765
|
0
|
|
|
|
|
0
|
$self->debug("Consed.pm : \$self is $self\n"); |
|
766
|
0
|
|
|
|
|
0
|
my (@points,$trimmed_sequence); |
|
767
|
0
|
0
|
|
|
|
0
|
if (!$self->{'doublets_set'}) { |
|
768
|
0
|
|
|
|
|
0
|
$self->debug("You need to set the doublets before you use ". |
|
769
|
|
|
|
|
|
|
"set_trim_points_singlets_and_doublets. Doing that now."); |
|
770
|
0
|
|
|
|
|
0
|
$self->set_doublets(); |
|
771
|
|
|
|
|
|
|
} |
|
772
|
0
|
|
|
|
|
0
|
foreach (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
0
|
|
|
773
|
0
|
0
|
|
|
|
0
|
if ($self->{'contigs'}->{$_}->{'class'} eq "singlet") { |
|
774
|
0
|
|
|
|
|
0
|
$self->debug("Singlet $_\n"); |
|
775
|
|
|
|
|
|
|
# this is what Warehouse wants |
|
776
|
|
|
|
|
|
|
# my ($self,$sequence,$quality,$name) = @_; |
|
777
|
|
|
|
|
|
|
# this is what Bio::Tools::Alignment::Trim::trim_singlet wants: |
|
778
|
|
|
|
|
|
|
# my ($self,$sequence,$quality,$name,$class) = @_; |
|
779
|
|
|
|
|
|
|
# the following several lines are to make the parameter passing legible. |
|
780
|
0
|
|
|
|
|
0
|
my ($sequence,$quality,$name,$class); |
|
781
|
0
|
|
|
|
|
0
|
$sequence = $self->{'contigs'}->{$_}->{'consensus'}; |
|
782
|
0
|
0
|
|
|
|
0
|
if (!$self->{'contigs'}->{$_}->{'quality'}) { $quality = "unset"; } |
|
|
0
|
|
|
|
|
0
|
|
|
783
|
0
|
|
|
|
|
0
|
else { $quality = $self->{'contigs'}->{$_}->{'quality'}; } |
|
784
|
0
|
|
|
|
|
0
|
$name = $self->{'contigs'}->{$_}->{'name'}; |
|
785
|
0
|
|
|
|
|
0
|
$class = $self->{'contigs'}->{$_}->{'class'}; |
|
786
|
0
|
|
|
|
|
0
|
@points = @{$self->{'o_trim'}->trim_singlet($sequence,$quality,$name,$class)}; |
|
|
0
|
|
|
|
|
0
|
|
|
787
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$_}->{'start_point'} = $points[0]; |
|
788
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$_}->{'end_point'} = $points[1]; |
|
789
|
|
|
|
|
|
|
$self->{'contigs'}->{$_}->{'sequence_trimmed'} = |
|
790
|
0
|
|
|
|
|
0
|
substr($self->{contigs}->{$_}->{'consensus'},$points[0],$points[1]-$points[0]); |
|
791
|
|
|
|
|
|
|
} |
|
792
|
|
|
|
|
|
|
} |
|
793
|
0
|
|
|
|
|
0
|
$self->debug("Bio::Tools::Alignment::Consed::set_trim_points_singlets". |
|
794
|
|
|
|
|
|
|
"_and_singletons: Done setting the quality trimpoints.\n"); |
|
795
|
0
|
|
|
|
|
0
|
return; |
|
796
|
|
|
|
|
|
|
} # end set_trim_points_singlet |
|
797
|
|
|
|
|
|
|
|
|
798
|
|
|
|
|
|
|
=head2 set_trim_points_doublets() |
|
799
|
|
|
|
|
|
|
|
|
800
|
|
|
|
|
|
|
Title : set_trim_points_doublets() |
|
801
|
|
|
|
|
|
|
Usage : $o_consed->set_trim_points_doublets(); |
|
802
|
|
|
|
|
|
|
Function: Set the trim points for doublets based on quality. Uses the |
|
803
|
|
|
|
|
|
|
Bio::Tools::Alignment::Trim object. Use at your own risk because |
|
804
|
|
|
|
|
|
|
the Bio::Tools::Alignment::Trim object was designed specifically |
|
805
|
|
|
|
|
|
|
for me and is mysterious in its ways. Every time somebody other |
|
806
|
|
|
|
|
|
|
then me uses it you risk a biblical plague being loosed on your |
|
807
|
|
|
|
|
|
|
city. |
|
808
|
|
|
|
|
|
|
Returns : Nothing. |
|
809
|
|
|
|
|
|
|
Args : None. |
|
810
|
|
|
|
|
|
|
Notes : Working on exceptions here. |
|
811
|
|
|
|
|
|
|
|
|
812
|
|
|
|
|
|
|
See L for more information |
|
813
|
|
|
|
|
|
|
|
|
814
|
|
|
|
|
|
|
=cut |
|
815
|
|
|
|
|
|
|
|
|
816
|
|
|
|
|
|
|
sub set_trim_points_doublets { |
|
817
|
0
|
|
|
0
|
1
|
0
|
my $self = shift; |
|
818
|
0
|
|
|
|
|
0
|
my @points; |
|
819
|
0
|
|
|
|
|
0
|
$self->debug("Bio::Tools::Alignment::Consed::set_trim_points_doublets:". |
|
820
|
|
|
|
|
|
|
" Restoring zeros for doublets.\n"); |
|
821
|
|
|
|
|
|
|
# &show_missing_sequence($self); |
|
822
|
0
|
|
|
|
|
0
|
$self->debug("Bio::Tools::Alignment::Consed::set_trim_points_doublets:". |
|
823
|
|
|
|
|
|
|
" Setting doublet trim points.\n"); |
|
824
|
0
|
|
|
|
|
0
|
foreach (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
0
|
|
|
825
|
0
|
0
|
|
|
|
0
|
if ($self->{'contigs'}->{$_}->{'class'} eq "doublet") { |
|
826
|
|
|
|
|
|
|
# my ($self,$sequence,$quality,$name,$class) = @_; |
|
827
|
0
|
|
|
|
|
0
|
my @quals = split(' ',$self->{'contigs'}->{$_}->{'quality'}); |
|
828
|
|
|
|
|
|
|
|
|
829
|
|
|
|
|
|
|
@points = $self->{o_trim}->trim_doublet |
|
830
|
|
|
|
|
|
|
($self->{'contigs'}->{$_}->{'consensus'}, |
|
831
|
|
|
|
|
|
|
$self->{'contigs'}->{$_}->{'quality'}, |
|
832
|
|
|
|
|
|
|
$self->{'contigs'}->{$_}->{name}, |
|
833
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$_}->{'class'}); |
|
834
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$_}->{'start_point'} = $points[0]; |
|
835
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$_}->{'end_point'} = $points[1]; |
|
836
|
|
|
|
|
|
|
# now set this |
|
837
|
|
|
|
|
|
|
$self->{'contigs'}->{$_}->{'sequence_trimmed'} = |
|
838
|
0
|
|
|
|
|
0
|
substr($self->{contigs}->{$_}->{'consensus'}, |
|
839
|
|
|
|
|
|
|
$points[0],$points[1]-$points[0]); |
|
840
|
|
|
|
|
|
|
# 010102 the deprecated way to do things: |
|
841
|
|
|
|
|
|
|
} |
|
842
|
|
|
|
|
|
|
} |
|
843
|
0
|
|
|
|
|
0
|
$self->debug("Bio::Tools::Alignment::Consed::set_trim_points_doublets:". |
|
844
|
|
|
|
|
|
|
" Done setting doublet trim points.\n"); |
|
845
|
0
|
|
|
|
|
0
|
return; |
|
846
|
|
|
|
|
|
|
} # end set_trim_points_doublets |
|
847
|
|
|
|
|
|
|
|
|
848
|
|
|
|
|
|
|
=head2 get_trimmed_sequence_by_name($name) |
|
849
|
|
|
|
|
|
|
|
|
850
|
|
|
|
|
|
|
Title : get_trimmed_sequence_by_name($name) |
|
851
|
|
|
|
|
|
|
Usage : $o_consed->get_trimmed_sequence_by_name($name); |
|
852
|
|
|
|
|
|
|
Function: Returns the trimmed_sequence of a contig with {name} eq $name. |
|
853
|
|
|
|
|
|
|
Returns : A scalar- the trimmed sequence. |
|
854
|
|
|
|
|
|
|
Args : The {name} of a contig. |
|
855
|
|
|
|
|
|
|
Notes : |
|
856
|
|
|
|
|
|
|
|
|
857
|
|
|
|
|
|
|
=cut |
|
858
|
|
|
|
|
|
|
|
|
859
|
|
|
|
|
|
|
sub get_trimmed_sequence_by_name { |
|
860
|
0
|
|
|
0
|
1
|
0
|
my ($self,$name) = @_; |
|
861
|
0
|
|
|
|
|
0
|
my $trimmed_sequence; |
|
862
|
0
|
|
|
|
|
0
|
my $contigname = &get_contig_number_by_name($self,$name); |
|
863
|
0
|
|
|
|
|
0
|
my $class = $self->{'contigs'}->{$contigname}->{'class'}; |
|
864
|
|
|
|
|
|
|
# what is this business and who was smoking crack while writing this? |
|
865
|
|
|
|
|
|
|
# if ($class eq "singlet") { |
|
866
|
|
|
|
|
|
|
# send the sequence, the quality, and the name |
|
867
|
|
|
|
|
|
|
# $trimmed_sequence = $self->{o_trim}->trim_singlet |
|
868
|
|
|
|
|
|
|
# ($self->{'contigs'}->{$contigname}->{consensus}, |
|
869
|
|
|
|
|
|
|
# $self->{'contigs'}->{$contigname}->{'quality'},$name); |
|
870
|
|
|
|
|
|
|
# } |
|
871
|
0
|
|
|
|
|
0
|
return $self->{'contigs'}->{$contigname}->{'sequence_trimmed'}; |
|
872
|
|
|
|
|
|
|
} |
|
873
|
|
|
|
|
|
|
|
|
874
|
|
|
|
|
|
|
=head2 set_dash_present_in_sequence_name("yes") |
|
875
|
|
|
|
|
|
|
|
|
876
|
|
|
|
|
|
|
Title : set_dash_present_in_sequence_name("yes") |
|
877
|
|
|
|
|
|
|
Usage : $o_consed->set_dash_present_in_sequence_name("yes"); |
|
878
|
|
|
|
|
|
|
Function: Deprecated. Part of an uncompleted thought. ("Oooh! Shiny!") |
|
879
|
|
|
|
|
|
|
Returns : Nothing. |
|
880
|
|
|
|
|
|
|
Args : "yes" to set {dash_present_in_sequence_name} to 1 |
|
881
|
|
|
|
|
|
|
Notes : |
|
882
|
|
|
|
|
|
|
|
|
883
|
|
|
|
|
|
|
=cut |
|
884
|
|
|
|
|
|
|
|
|
885
|
|
|
|
|
|
|
sub set_dash_present_in_sequence_name { |
|
886
|
0
|
|
|
0
|
1
|
0
|
my ($self,$dash_present) = @_; |
|
887
|
0
|
0
|
|
|
|
0
|
if ($dash_present eq "yes") { |
|
888
|
0
|
|
|
|
|
0
|
$self->{'dash_present_in_sequence_name'} = 1; |
|
889
|
|
|
|
|
|
|
} |
|
890
|
|
|
|
|
|
|
else { |
|
891
|
0
|
|
|
|
|
0
|
$self->{'dash_present_in_sequence_name'} = 0; |
|
892
|
|
|
|
|
|
|
} |
|
893
|
0
|
|
|
|
|
0
|
return $self->{'dash_present_in_sequence_name'}; |
|
894
|
|
|
|
|
|
|
} # end set_dash_present_in_sequence_name |
|
895
|
|
|
|
|
|
|
|
|
896
|
|
|
|
|
|
|
=head2 set_doublets() |
|
897
|
|
|
|
|
|
|
|
|
898
|
|
|
|
|
|
|
Title : set_doublets() |
|
899
|
|
|
|
|
|
|
Usage : $o_consed->set_doublets(); |
|
900
|
|
|
|
|
|
|
Function: Find pairs that have similar names and mark them as doublets |
|
901
|
|
|
|
|
|
|
and set the {name}. |
|
902
|
|
|
|
|
|
|
Returns : 0 or 1. |
|
903
|
|
|
|
|
|
|
Args : None. |
|
904
|
|
|
|
|
|
|
|
|
905
|
|
|
|
|
|
|
A complicated subroutine that iterates over the |
|
906
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed looking for contigs of 2. If the forward |
|
907
|
|
|
|
|
|
|
and reverse designator are removed from each of the reads in |
|
908
|
|
|
|
|
|
|
{'member_array'} and the remaining reads are the same, {name} is set |
|
909
|
|
|
|
|
|
|
to that name and the contig's class is set as "doublet". If any of |
|
910
|
|
|
|
|
|
|
those cases fail the contig is marked as a "pair". |
|
911
|
|
|
|
|
|
|
|
|
912
|
|
|
|
|
|
|
=cut |
|
913
|
|
|
|
|
|
|
|
|
914
|
|
|
|
|
|
|
#' make my emacs happy |
|
915
|
|
|
|
|
|
|
|
|
916
|
|
|
|
|
|
|
sub set_doublets { |
|
917
|
1
|
|
|
1
|
1
|
477
|
my ($self) = @_; |
|
918
|
|
|
|
|
|
|
# set the designators in the Bio::Tools::Alignment::Trim object |
|
919
|
|
|
|
|
|
|
|
|
920
|
|
|
|
|
|
|
$self->{'o_trim'}->set_designators($self->{'reverse_designator'}, |
|
921
|
1
|
|
|
|
|
8
|
$self->{'forward_designator'}); |
|
922
|
1
|
|
|
|
|
2
|
foreach my $key_contig (sort keys %{$self->{'contigs'}}) { |
|
|
1
|
|
|
|
|
54
|
|
|
923
|
|
|
|
|
|
|
|
|
924
|
|
|
|
|
|
|
# if there is a member array (why would there not be? This should be a die()able offence |
|
925
|
|
|
|
|
|
|
# but for now I will leave it |
|
926
|
118
|
50
|
|
|
|
167
|
if ($self->{'contigs'}->{$key_contig}->{'member_array'}) { |
|
927
|
|
|
|
|
|
|
# if there are two reads in this contig |
|
928
|
|
|
|
|
|
|
# i am pretty sure that this is wrong but i am keeping it for reference |
|
929
|
|
|
|
|
|
|
# if (@{$self->{'contigs'}->{$key_contig}->{'member_array'}} == 2 || !$self->{'contigs'}->{$key_contig}->{'class'}) { |
|
930
|
|
|
|
|
|
|
# |
|
931
|
|
|
|
|
|
|
# WRONG. Was I on crack? |
|
932
|
118
|
100
|
|
|
|
64
|
if (@{$self->{'contigs'}->{$key_contig}->{'member_array'}} == 2) { |
|
|
118
|
100
|
|
|
|
136
|
|
|
|
|
50
|
|
|
|
|
|
|
933
|
46
|
|
|
|
|
37
|
$self->{'contigs'}->{$key_contig}->{'num_members'} = 2; |
|
934
|
46
|
|
|
|
|
61
|
$self->debug("\tThere are 2 members! Looking for the contig name...\n"); |
|
935
|
46
|
|
|
|
|
63
|
my $name = _get_contig_name($self,$self->{'contigs'}->{$key_contig}->{'member_array'}); |
|
936
|
46
|
100
|
|
|
|
114
|
$self->debug("The name is $name\n") if defined $name; |
|
937
|
46
|
100
|
|
|
|
47
|
if ($name) { |
|
938
|
45
|
|
|
|
|
48
|
$self->{'contigs'}->{$key_contig}->{'name'} = $name; |
|
939
|
45
|
|
|
|
|
52
|
$self->{'contigs'}->{$key_contig}->{'class'} = "doublet"; |
|
940
|
|
|
|
|
|
|
} else { |
|
941
|
1
|
|
|
|
|
4
|
$self->debug("$key_contig is a pair.\n"); |
|
942
|
1
|
|
|
|
|
2
|
$self->{'contigs'}->{$key_contig}->{'class'} = "pair"; |
|
943
|
|
|
|
|
|
|
} |
|
944
|
|
|
|
|
|
|
} |
|
945
|
|
|
|
|
|
|
# this is all fair and good but what about singlets? |
|
946
|
|
|
|
|
|
|
# they have one reads in the member_array but certainly are not singletons |
|
947
|
72
|
|
|
|
|
75
|
elsif (@{$self->{'contigs'}->{$key_contig}->{'member_array'}} == 1) { |
|
948
|
|
|
|
|
|
|
# set the name to be the name of the read |
|
949
|
68
|
|
|
|
|
32
|
$self->{'contigs'}->{$key_contig}->{name} = @{$self->{'contigs'}->{$key_contig}->{'member_array'}}[0]; |
|
|
68
|
|
|
|
|
83
|
|
|
950
|
|
|
|
|
|
|
# set the number of members to be one |
|
951
|
68
|
|
|
|
|
51
|
$self->{'contigs'}->{$key_contig}->{num_members} = 1; |
|
952
|
|
|
|
|
|
|
# if this was a singlet, it would already belong to the class "singlet" |
|
953
|
|
|
|
|
|
|
# so leave it alone |
|
954
|
|
|
|
|
|
|
# if it is not a singlet, it is a singleton! lablel it appropriately |
|
955
|
68
|
100
|
|
|
|
75
|
unless ($self->{'contigs'}->{$key_contig}->{'class'}) { |
|
956
|
3
|
|
|
|
|
4
|
$self->{'contigs'}->{$key_contig}->{'class'} = "singleton"; |
|
957
|
|
|
|
|
|
|
} |
|
958
|
|
|
|
|
|
|
} |
|
959
|
|
|
|
|
|
|
# set the multiplet characteristics |
|
960
|
4
|
|
|
|
|
9
|
elsif (@{$self->{'contigs'}->{$key_contig}->{'member_array'}} >= 3) { |
|
961
|
4
|
|
|
|
|
5
|
$self->{'contigs'}->{$key_contig}->{'num_members'} = @{$self->{'contigs'}->{$key_contig}->{'member_array'}}; |
|
|
4
|
|
|
|
|
8
|
|
|
962
|
4
|
|
|
|
|
6
|
$self->{'contigs'}->{$key_contig}->{'class'} = "multiplet"; |
|
963
|
|
|
|
|
|
|
} |
|
964
|
118
|
|
|
|
|
85
|
$self->{'contigs'}->{$key_contig}->{'num_members'} = @{$self->{'contigs'}->{$key_contig}->{'member_array'}}; |
|
|
118
|
|
|
|
|
135
|
|
|
965
|
|
|
|
|
|
|
|
|
966
|
|
|
|
|
|
|
} |
|
967
|
|
|
|
|
|
|
} |
|
968
|
1
|
|
|
|
|
15
|
$self->{'doublets_set'} = "done"; |
|
969
|
1
|
|
|
|
|
7
|
return 0; |
|
970
|
|
|
|
|
|
|
} # end set_doublets |
|
971
|
|
|
|
|
|
|
|
|
972
|
|
|
|
|
|
|
=head2 set_singlets |
|
973
|
|
|
|
|
|
|
|
|
974
|
|
|
|
|
|
|
Title : set_singlets |
|
975
|
|
|
|
|
|
|
Usage : $o_consed->set_singlets(); |
|
976
|
|
|
|
|
|
|
Function: Read in a singlets file and place them into the |
|
977
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. |
|
978
|
|
|
|
|
|
|
Returns : Nothing. |
|
979
|
|
|
|
|
|
|
Args : A scalar to turn on verbose parsing of the singlets file. |
|
980
|
|
|
|
|
|
|
Notes : |
|
981
|
|
|
|
|
|
|
|
|
982
|
|
|
|
|
|
|
=cut |
|
983
|
|
|
|
|
|
|
|
|
984
|
|
|
|
|
|
|
sub set_singlets { |
|
985
|
|
|
|
|
|
|
# parse out the contents of the singlets file |
|
986
|
1
|
|
|
1
|
1
|
365
|
my ($self) = @_; |
|
987
|
1
|
|
|
|
|
8
|
$self->debug("Bio::Tools::Alignment::Consed Adding singlets to the contig hash...\n"); |
|
988
|
1
|
|
|
|
|
2
|
my $full_filename = $self->{'filename'}; |
|
989
|
1
|
|
|
|
|
5
|
$self->debug("Bio::Tools::Alignment::Consed::set_singlets: \$full_filename is $full_filename\n"); |
|
990
|
1
|
50
|
|
|
|
19
|
$full_filename =~ s#\\#\/#g if $^O =~ m/mswin/i; |
|
991
|
1
|
|
|
|
|
5
|
$full_filename =~ m/(.*\/)(.*ace.*)$/; |
|
992
|
1
|
|
|
|
|
3
|
my ($base_path,$filename) = ($1,$2); |
|
993
|
1
|
|
|
|
|
6
|
$self->debug("Bio::Tools::Alignment::Consed::set_singlets: singlets filename is $filename and \$base_path is $base_path\n"); |
|
994
|
1
|
|
|
|
|
3
|
$filename =~ m/(.*)ace.*$/; |
|
995
|
1
|
|
|
|
|
3
|
my $singletsfile = $base_path.$1."singlets"; |
|
996
|
1
|
|
|
|
|
6
|
$self->debug("\$singletsfile is $singletsfile\n"); |
|
997
|
1
|
50
|
|
|
|
37
|
if (!-f $singletsfile) { |
|
998
|
|
|
|
|
|
|
# there is no singlets file. |
|
999
|
0
|
|
|
|
|
0
|
$self->{'singlets_set'} = "done"; |
|
1000
|
0
|
|
|
|
|
0
|
return; |
|
1001
|
|
|
|
|
|
|
} |
|
1002
|
1
|
|
|
|
|
3
|
$self->debug("$singletsfile is indeed a file. Trying to open it...\n"); |
|
1003
|
1
|
|
|
|
|
5
|
my $singlets_fh = Bio::Root::IO->new(-file => $singletsfile); |
|
1004
|
1
|
|
|
|
|
2
|
my ($sequence,$name,$count); |
|
1005
|
1
|
|
|
|
|
3
|
while ($_ = $singlets_fh->_readline()) { |
|
1006
|
1291
|
|
|
|
|
913
|
chomp $_; |
|
1007
|
1291
|
100
|
|
|
|
1407
|
if (/\>/) { |
|
1008
|
65
|
100
|
66
|
|
|
158
|
if ($name && $sequence) { |
|
1009
|
64
|
|
|
|
|
271
|
$self->debug("Adding $name with sequence $sequence to hash...\n"); |
|
1010
|
64
|
|
|
|
|
69
|
push @{$self->{'contigs'}->{$name}->{'member_array'}},$name; |
|
|
64
|
|
|
|
|
266
|
|
|
1011
|
64
|
|
|
|
|
86
|
$self->{'contigs'}->{$name}->{'consensus'} = $sequence; |
|
1012
|
64
|
|
|
|
|
75
|
$self->{'contigs'}->{$name}->{'name'} = $name; |
|
1013
|
64
|
|
|
|
|
59
|
$self->{'contigs'}->{$name}->{"singlet"} = 1; |
|
1014
|
64
|
|
|
|
|
64
|
$self->{'contigs'}->{$name}->{'class'} = "singlet"; |
|
1015
|
|
|
|
|
|
|
} |
|
1016
|
65
|
|
|
|
|
54
|
$sequence = $name = undef; |
|
1017
|
65
|
|
|
|
|
40
|
$count++; |
|
1018
|
65
|
|
|
|
|
327
|
m/^\>(.*)\s\sCHROMAT/; |
|
1019
|
65
|
|
|
|
|
92
|
$name = $1; |
|
1020
|
65
|
50
|
|
|
|
147
|
if (!$name) { |
|
1021
|
0
|
|
|
|
|
0
|
m/\>(\S+)\s/; |
|
1022
|
0
|
|
|
|
|
0
|
$name = $1; |
|
1023
|
|
|
|
|
|
|
} |
|
1024
|
|
|
|
|
|
|
} |
|
1025
|
1226
|
|
|
|
|
1821
|
else { $sequence .= $_; } |
|
1026
|
|
|
|
|
|
|
} |
|
1027
|
1
|
50
|
33
|
|
|
7
|
if ($name && $sequence) { |
|
1028
|
1
|
|
|
|
|
6
|
$self->debug("Pushing the last of the singlets ($name)\n"); |
|
1029
|
1
|
|
|
|
|
2
|
@{$self->{'contigs'}->{$name}->{'member_array'}} = $name; |
|
|
1
|
|
|
|
|
3
|
|
|
1030
|
1
|
|
|
|
|
5
|
$self->{'contigs'}->{$name}->{'consensus'} = $sequence; |
|
1031
|
1
|
|
|
|
|
5
|
$self->{'contigs'}->{$name}->{'name'} = $name; |
|
1032
|
1
|
|
|
|
|
2
|
$self->{'contigs'}->{$name}->{"singlet"} = 1; |
|
1033
|
1
|
|
|
|
|
2
|
$self->{'contigs'}->{$name}->{'class'} = "singlet"; |
|
1034
|
|
|
|
|
|
|
} |
|
1035
|
1
|
|
|
|
|
3
|
$self->debug("Bio::Tools::Alignment::Consed::set_singlets: Done adding singlets to the singlets hash.\n"); |
|
1036
|
1
|
|
|
|
|
2
|
$self->{'singlets_set'} = "done"; |
|
1037
|
1
|
|
|
|
|
17
|
return 0; |
|
1038
|
|
|
|
|
|
|
} # end sub set_singlets |
|
1039
|
|
|
|
|
|
|
|
|
1040
|
|
|
|
|
|
|
=head2 get_singlets() |
|
1041
|
|
|
|
|
|
|
|
|
1042
|
|
|
|
|
|
|
Title : get_singlets() |
|
1043
|
|
|
|
|
|
|
Usage : $o_consed->get_singlets(); |
|
1044
|
|
|
|
|
|
|
Function: Return the keynames of the singlets. |
|
1045
|
|
|
|
|
|
|
Returns : An array containing the keynames of all |
|
1046
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed sequences in the class "singlet". |
|
1047
|
|
|
|
|
|
|
Args : None. |
|
1048
|
|
|
|
|
|
|
Notes : |
|
1049
|
|
|
|
|
|
|
|
|
1050
|
|
|
|
|
|
|
=cut |
|
1051
|
|
|
|
|
|
|
|
|
1052
|
|
|
|
|
|
|
sub get_singlets { |
|
1053
|
|
|
|
|
|
|
# returns an array of singlet names |
|
1054
|
|
|
|
|
|
|
# singlets have "singlet"=1 in the hash |
|
1055
|
3
|
|
|
3
|
1
|
5
|
my $self = shift; |
|
1056
|
3
|
50
|
|
|
|
7
|
if (!$self->{singlets_set}) { |
|
1057
|
0
|
|
|
|
|
0
|
$self->debug("You need to set the singlets before you get them. Doing that now."); |
|
1058
|
0
|
|
|
|
|
0
|
$self->set_singlets(); |
|
1059
|
|
|
|
|
|
|
} |
|
1060
|
|
|
|
|
|
|
|
|
1061
|
3
|
|
|
|
|
4
|
my (@singlets,@array); |
|
1062
|
3
|
|
|
|
|
4
|
foreach my $key (sort keys %{$self->{'contigs'}}) { |
|
|
3
|
|
|
|
|
138
|
|
|
1063
|
|
|
|
|
|
|
# @array = @{$Consed::contigs{$key}->{'member_array'}}; |
|
1064
|
|
|
|
|
|
|
# somethimes a user will try to get a list of singlets before the classes for the rest of the |
|
1065
|
|
|
|
|
|
|
# contigs has been set (see t/test.t for how I figured this out. |
|
1066
|
|
|
|
|
|
|
# so either way, just return class=singlets |
|
1067
|
354
|
100
|
|
|
|
608
|
if (!$self->{'contigs'}->{$key}->{'class'}) { |
|
|
|
100
|
|
|
|
|
|
|
1068
|
|
|
|
|
|
|
# print("$key has no class. why?\n"); |
|
1069
|
|
|
|
|
|
|
} |
|
1070
|
|
|
|
|
|
|
elsif ($self->{'contigs'}->{$key}->{'class'} eq "singlet") { |
|
1071
|
195
|
|
|
|
|
153
|
push @singlets,$key; |
|
1072
|
|
|
|
|
|
|
} |
|
1073
|
|
|
|
|
|
|
} |
|
1074
|
3
|
|
|
|
|
26
|
return @singlets; |
|
1075
|
|
|
|
|
|
|
} |
|
1076
|
|
|
|
|
|
|
|
|
1077
|
|
|
|
|
|
|
=head2 set_quality_by_name($name,$quality) |
|
1078
|
|
|
|
|
|
|
|
|
1079
|
|
|
|
|
|
|
Title : set_quality_by_name($name,$quality) |
|
1080
|
|
|
|
|
|
|
Usage : $o_consed->set_quality_by_name($name,$quality); |
|
1081
|
|
|
|
|
|
|
Function: Deprecated. Make the contig with {name} have {'quality'} $quality. |
|
1082
|
|
|
|
|
|
|
Probably used for testing. |
|
1083
|
|
|
|
|
|
|
Returns : Nothing. |
|
1084
|
|
|
|
|
|
|
Args : The name of a contig and a scalar for its quality. |
|
1085
|
|
|
|
|
|
|
Notes : Deprecated. |
|
1086
|
|
|
|
|
|
|
|
|
1087
|
|
|
|
|
|
|
=cut |
|
1088
|
|
|
|
|
|
|
|
|
1089
|
|
|
|
|
|
|
sub set_quality_by_name { |
|
1090
|
|
|
|
|
|
|
# this is likely deprecated |
|
1091
|
0
|
|
|
0
|
1
|
0
|
my ($self,$name,$quality) = shift; |
|
1092
|
0
|
|
|
|
|
0
|
my $return; |
|
1093
|
0
|
|
|
|
|
0
|
foreach (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
0
|
|
|
1094
|
0
|
0
|
0
|
|
|
0
|
if ($self->{'contigs'} eq "$name" || $self->{'contigs'}->{'name'} eq "$name") { |
|
1095
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{'quality'} = $quality; |
|
1096
|
0
|
|
|
|
|
0
|
$return=1; |
|
1097
|
|
|
|
|
|
|
} |
|
1098
|
|
|
|
|
|
|
} |
|
1099
|
0
|
0
|
|
|
|
0
|
if ($return) { return "0"; } else { return "1"; } |
|
|
0
|
|
|
|
|
0
|
|
|
|
0
|
|
|
|
|
0
|
|
|
1100
|
|
|
|
|
|
|
} # end set quality by name |
|
1101
|
|
|
|
|
|
|
|
|
1102
|
|
|
|
|
|
|
=head2 set_singlet_quality() |
|
1103
|
|
|
|
|
|
|
|
|
1104
|
|
|
|
|
|
|
Title : set_singlet_quality() |
|
1105
|
|
|
|
|
|
|
Usage : $o_consed->set_singlet_quality(); |
|
1106
|
|
|
|
|
|
|
Function: For each singlet, go to the appropriate file in phd_dir and read |
|
1107
|
|
|
|
|
|
|
in the phred quality for that read and place it into {'quality'} |
|
1108
|
|
|
|
|
|
|
Returns : 0 or 1. |
|
1109
|
|
|
|
|
|
|
Args : None. |
|
1110
|
|
|
|
|
|
|
Notes : This is the next subroutine that will receive substantial revision |
|
1111
|
|
|
|
|
|
|
in the next little while. It really should eval the creation of |
|
1112
|
|
|
|
|
|
|
Bio::Tools::Alignment::Phred objects and go from there. |
|
1113
|
|
|
|
|
|
|
|
|
1114
|
|
|
|
|
|
|
=cut |
|
1115
|
|
|
|
|
|
|
|
|
1116
|
|
|
|
|
|
|
sub set_singlet_quality { |
|
1117
|
0
|
|
|
0
|
1
|
0
|
my $self = shift; |
|
1118
|
0
|
|
|
|
|
0
|
my $full_filename = $self->{'filename'}; |
|
1119
|
0
|
0
|
|
|
|
0
|
$full_filename =~ s#\\#\/#g if $^O =~ m/mswin/i; |
|
1120
|
0
|
|
|
|
|
0
|
$full_filename =~ m/(.*\/)(.*)ace.*$/; |
|
1121
|
0
|
|
|
|
|
0
|
my ($base_path,$filename) = ($1,"$2"."qual"); |
|
1122
|
0
|
|
|
|
|
0
|
my $singletsfile = $base_path.$filename; |
|
1123
|
0
|
0
|
|
|
|
0
|
if (-f $singletsfile) { |
|
1124
|
|
|
|
|
|
|
# print("$singletsfile is indeed a file. Trying to open it...\n"); |
|
1125
|
|
|
|
|
|
|
} |
|
1126
|
|
|
|
|
|
|
else { |
|
1127
|
0
|
|
|
|
|
0
|
$self->warn("$singletsfile is not a file. Sorry.\n"); |
|
1128
|
0
|
|
|
|
|
0
|
return; |
|
1129
|
|
|
|
|
|
|
} |
|
1130
|
0
|
|
|
|
|
0
|
my $singlets_fh = Bio::Root::IO->new(-file => $singletsfile); |
|
1131
|
0
|
|
|
|
|
0
|
my ($sequence,$name,$count); |
|
1132
|
0
|
|
|
|
|
0
|
my ($identity,$line,$quality,@qline); |
|
1133
|
0
|
|
|
|
|
0
|
while ($line = $singlets_fh->_readline()) { |
|
1134
|
0
|
|
|
|
|
0
|
chomp $line; |
|
1135
|
0
|
0
|
|
|
|
0
|
if ($line =~ /^\>/) { |
|
1136
|
0
|
|
|
|
|
0
|
$quality = undef; |
|
1137
|
0
|
|
|
|
|
0
|
$line =~ m/\>(\S*)\s/; |
|
1138
|
0
|
|
|
|
|
0
|
$identity = $1; |
|
1139
|
|
|
|
|
|
|
} |
|
1140
|
|
|
|
|
|
|
else { |
|
1141
|
0
|
0
|
|
|
|
0
|
if ($self->{'contigs'}->{$identity}) { |
|
1142
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$identity}->{'quality'} .= "$line "; |
|
1143
|
|
|
|
|
|
|
} |
|
1144
|
|
|
|
|
|
|
} |
|
1145
|
|
|
|
|
|
|
|
|
1146
|
|
|
|
|
|
|
} |
|
1147
|
0
|
|
|
|
|
0
|
return 0; |
|
1148
|
|
|
|
|
|
|
} |
|
1149
|
|
|
|
|
|
|
|
|
1150
|
|
|
|
|
|
|
=head2 set_contig_quality() |
|
1151
|
|
|
|
|
|
|
|
|
1152
|
|
|
|
|
|
|
Title : set_contig_quality() |
|
1153
|
|
|
|
|
|
|
Usage : $o_consed->set_contig_quality(); |
|
1154
|
|
|
|
|
|
|
Function: Deprecated. |
|
1155
|
|
|
|
|
|
|
Returns : Deprecated. |
|
1156
|
|
|
|
|
|
|
Args : Deprecated. |
|
1157
|
|
|
|
|
|
|
Notes : Deprecated. Really. Trust me. |
|
1158
|
|
|
|
|
|
|
|
|
1159
|
|
|
|
|
|
|
=cut |
|
1160
|
|
|
|
|
|
|
|
|
1161
|
|
|
|
|
|
|
sub set_contig_quality { |
|
1162
|
|
|
|
|
|
|
# note: contigs _include_ singletons but _not_ singlets |
|
1163
|
0
|
|
|
0
|
1
|
0
|
my ($self) = shift; |
|
1164
|
|
|
|
|
|
|
# the unexpected results I am referring to here are a doubling of quality values. |
|
1165
|
|
|
|
|
|
|
# the profanity I uttered on discovering this reminded me of the simpsons: |
|
1166
|
|
|
|
|
|
|
# Ned Flanders: "That is the loudest profanity I have ever heard!" |
|
1167
|
0
|
|
|
|
|
0
|
$self->warn("set_contig_quality is deprecated and will likely produce unexpected results"); |
|
1168
|
0
|
|
|
|
|
0
|
my $full_filename = $self->{'filename'}; |
|
1169
|
|
|
|
|
|
|
# Run_SRC3700_2000-08-01_73+74.fasta.screen.contigs.qual |
|
1170
|
|
|
|
|
|
|
# from Consed.pm |
|
1171
|
0
|
0
|
|
|
|
0
|
$full_filename =~ s#\\#\/#g if $^O =~ m/mswin/i; |
|
1172
|
0
|
|
|
|
|
0
|
$full_filename =~ m/(.*\/)(.*)ace.*$/; |
|
1173
|
0
|
|
|
|
|
0
|
my ($base_path,$filename) = ($1,"$2"."contigs.qual"); |
|
1174
|
0
|
|
|
|
|
0
|
my $singletsfile = $base_path.$filename; |
|
1175
|
0
|
0
|
|
|
|
0
|
if (-f $singletsfile) { |
|
1176
|
|
|
|
|
|
|
# print("$singletsfile is indeed a file. Trying to open it...\n"); |
|
1177
|
|
|
|
|
|
|
} |
|
1178
|
|
|
|
|
|
|
else { |
|
1179
|
0
|
|
|
|
|
0
|
$self->warn("Bio::Tools::Alignment::Consed::set_contig_quality $singletsfile is not a file. Sorry.\n"); |
|
1180
|
0
|
|
|
|
|
0
|
return; |
|
1181
|
|
|
|
|
|
|
} |
|
1182
|
0
|
|
|
|
|
0
|
my $contig_quality_fh = Bio::Root::IO->new(-file => $singletsfile); |
|
1183
|
|
|
|
|
|
|
|
|
1184
|
0
|
|
|
|
|
0
|
my ($sequence,$name,$count,$identity,$line,$quality); |
|
1185
|
0
|
|
|
|
|
0
|
while ($line = $contig_quality_fh->_readline()) { |
|
1186
|
0
|
|
|
|
|
0
|
chomp $line; |
|
1187
|
0
|
0
|
|
|
|
0
|
if ($line =~ /^\>/) { |
|
1188
|
0
|
|
|
|
|
0
|
$quality = undef; |
|
1189
|
0
|
|
|
|
|
0
|
$line =~ m/\>.*Contig(\d+)\s/; |
|
1190
|
0
|
|
|
|
|
0
|
$identity = $1; |
|
1191
|
|
|
|
|
|
|
} |
|
1192
|
|
|
|
|
|
|
else { |
|
1193
|
0
|
0
|
|
|
|
0
|
if ($self->{'contigs'}->{$identity} ) { |
|
1194
|
0
|
|
|
|
|
0
|
$self->{'contigs'}->{$identity}->{'quality'} .= " $line"; |
|
1195
|
|
|
|
|
|
|
} |
|
1196
|
|
|
|
|
|
|
} |
|
1197
|
|
|
|
|
|
|
} |
|
1198
|
|
|
|
|
|
|
} # end set_contig_quality |
|
1199
|
|
|
|
|
|
|
|
|
1200
|
|
|
|
|
|
|
=head2 get_multiplets() |
|
1201
|
|
|
|
|
|
|
|
|
1202
|
|
|
|
|
|
|
Title : get_multiplets() |
|
1203
|
|
|
|
|
|
|
Usage : $o_consed->get_multiplets(); |
|
1204
|
|
|
|
|
|
|
Function: Return the keynames of the multiplets. |
|
1205
|
|
|
|
|
|
|
Returns : Returns an array containing the keynames of all |
|
1206
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed sequences in the class "multiplet". |
|
1207
|
|
|
|
|
|
|
Args : None. |
|
1208
|
|
|
|
|
|
|
Notes : |
|
1209
|
|
|
|
|
|
|
|
|
1210
|
|
|
|
|
|
|
=cut |
|
1211
|
|
|
|
|
|
|
|
|
1212
|
|
|
|
|
|
|
sub get_multiplets { |
|
1213
|
|
|
|
|
|
|
# returns an array of multiplet names |
|
1214
|
|
|
|
|
|
|
# multiplets have # members > 2 |
|
1215
|
3
|
|
|
3
|
1
|
479
|
my $self = shift; |
|
1216
|
3
|
|
|
|
|
3
|
my (@multiplets,@array); |
|
1217
|
3
|
|
|
|
|
3
|
foreach my $key (sort keys %{$self->{'contigs'}}) { |
|
|
3
|
|
|
|
|
89
|
|
|
1218
|
354
|
50
|
|
|
|
399
|
if ($self->{'contigs'}->{$key}->{'class'}) { |
|
1219
|
354
|
100
|
|
|
|
427
|
if ($self->{'contigs'}->{$key}->{'class'} eq "multiplet") { |
|
1220
|
12
|
|
|
|
|
11
|
push @multiplets,$key; |
|
1221
|
|
|
|
|
|
|
} |
|
1222
|
|
|
|
|
|
|
} |
|
1223
|
|
|
|
|
|
|
} |
|
1224
|
3
|
|
|
|
|
17
|
return @multiplets; |
|
1225
|
|
|
|
|
|
|
} |
|
1226
|
|
|
|
|
|
|
|
|
1227
|
|
|
|
|
|
|
=head2 get_all_members() |
|
1228
|
|
|
|
|
|
|
|
|
1229
|
|
|
|
|
|
|
Title : get_all_members() |
|
1230
|
|
|
|
|
|
|
Usage : @all_members = $o_consed->get_all_members(); |
|
1231
|
|
|
|
|
|
|
Function: Return a list of all of the read names in the |
|
1232
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. |
|
1233
|
|
|
|
|
|
|
Returns : An array containing all of the elements in all of the |
|
1234
|
|
|
|
|
|
|
{'member_array'}s. |
|
1235
|
|
|
|
|
|
|
Args : None. |
|
1236
|
|
|
|
|
|
|
Notes : |
|
1237
|
|
|
|
|
|
|
|
|
1238
|
|
|
|
|
|
|
=cut |
|
1239
|
|
|
|
|
|
|
|
|
1240
|
|
|
|
|
|
|
sub get_all_members { |
|
1241
|
0
|
|
|
0
|
1
|
0
|
my $self = shift; |
|
1242
|
0
|
|
|
|
|
0
|
my @members; |
|
1243
|
0
|
|
|
|
|
0
|
foreach my $key (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
0
|
|
|
1244
|
0
|
0
|
|
|
|
0
|
if ($key =~ /^singlet/) { |
|
|
|
0
|
|
|
|
|
|
|
1245
|
0
|
|
|
|
|
0
|
push @members,$self->{'contigs'}->{$key}->{'member_array'}[0]; |
|
1246
|
|
|
|
|
|
|
} |
|
1247
|
|
|
|
|
|
|
elsif ($self->{'contigs'}->{$key}->{'member_array'}) { |
|
1248
|
0
|
|
|
|
|
0
|
push @members,@{$self->{'contigs'}->{$key}->{'member_array'}}; |
|
|
0
|
|
|
|
|
0
|
|
|
1249
|
|
|
|
|
|
|
} |
|
1250
|
|
|
|
|
|
|
# else { |
|
1251
|
|
|
|
|
|
|
# print("Bio::Tools::Alignment::Consed: $key is _not_ an array. Pushing $self->{'contigs'}->{$key}->{'member_array'} onto \@members\n"); |
|
1252
|
|
|
|
|
|
|
# push @members,$self->{'contigs'}->{$key}->{'member_array'}; |
|
1253
|
|
|
|
|
|
|
# } |
|
1254
|
|
|
|
|
|
|
} |
|
1255
|
0
|
|
|
|
|
0
|
return @members; |
|
1256
|
|
|
|
|
|
|
} |
|
1257
|
|
|
|
|
|
|
|
|
1258
|
|
|
|
|
|
|
=head2 sum_lets($total_only) |
|
1259
|
|
|
|
|
|
|
|
|
1260
|
|
|
|
|
|
|
Title : sum_lets($total_only) |
|
1261
|
|
|
|
|
|
|
Usage : $statistics = $o_consed->sum_lets($total_only); |
|
1262
|
|
|
|
|
|
|
Function: Provide numbers for how many sequences were accounted for in the |
|
1263
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. |
|
1264
|
|
|
|
|
|
|
Returns : If a scalar is present, returns the total number of |
|
1265
|
|
|
|
|
|
|
sequences accounted for in all classes. If no scalar passed |
|
1266
|
|
|
|
|
|
|
then returns a string that looks like this: |
|
1267
|
|
|
|
|
|
|
Singt/singn/doub/pair/mult/total : 2,0,1(2),0(0),0(0),4 |
|
1268
|
|
|
|
|
|
|
This example means the following: There were 1 singlets. |
|
1269
|
|
|
|
|
|
|
There were 0 singletons. There were 1 doublets for a total |
|
1270
|
|
|
|
|
|
|
of 2 sequences in this class. There were 0 pairs for a |
|
1271
|
|
|
|
|
|
|
total of 0 sequences in this class. There were 0 |
|
1272
|
|
|
|
|
|
|
multiplets for a total of 0 sequences in this class. There |
|
1273
|
|
|
|
|
|
|
were a total of 4 sequences accounted for in the |
|
1274
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed object. |
|
1275
|
|
|
|
|
|
|
Args : A scalar is optional to change the way the numbers are returned. |
|
1276
|
|
|
|
|
|
|
Notes: |
|
1277
|
|
|
|
|
|
|
|
|
1278
|
|
|
|
|
|
|
=cut |
|
1279
|
|
|
|
|
|
|
|
|
1280
|
|
|
|
|
|
|
sub sum_lets { |
|
1281
|
2
|
|
|
2
|
1
|
4
|
my ($self,$total_only) = @_; |
|
1282
|
2
|
|
|
|
|
1
|
my ($count,$count_multiplets,$multiplet_count); |
|
1283
|
2
|
|
|
|
|
5
|
my $singlets = &get_singlets($self); $count += $singlets; |
|
|
2
|
|
|
|
|
3
|
|
|
1284
|
2
|
|
|
|
|
4
|
my $doublets = &get_doublets($self); $count += ($doublets * 2); |
|
|
2
|
|
|
|
|
4
|
|
|
1285
|
2
|
|
|
|
|
6
|
my $pairs = &get_pairs($self); $count += ($pairs * 2); |
|
|
2
|
|
|
|
|
4
|
|
|
1286
|
2
|
|
|
|
|
4
|
my $singletons = &get_singletons($self); $count += $singletons; |
|
|
2
|
|
|
|
|
3
|
|
|
1287
|
2
|
|
|
|
|
4
|
my @multiplets = &get_multiplets($self); |
|
1288
|
2
|
|
|
|
|
3
|
$count_multiplets = @multiplets; |
|
1289
|
2
|
|
|
|
|
2
|
my $return_string; |
|
1290
|
2
|
|
|
|
|
3
|
foreach (@multiplets) { |
|
1291
|
8
|
|
|
|
|
8
|
my $number_members = $self->{'contigs'}->{$_}->{num_members}; |
|
1292
|
8
|
|
|
|
|
6
|
$multiplet_count += $number_members; |
|
1293
|
|
|
|
|
|
|
} |
|
1294
|
2
|
50
|
|
|
|
5
|
if ($multiplet_count) { |
|
1295
|
2
|
|
|
|
|
1
|
$count += $multiplet_count; |
|
1296
|
|
|
|
|
|
|
} |
|
1297
|
2
|
|
|
|
|
4
|
foreach (qw(multiplet_count singlets doublets pairs singletons |
|
1298
|
|
|
|
|
|
|
multiplets count_multiplets)) { |
|
1299
|
1
|
|
|
1
|
|
5
|
no strict 'refs'; # renege for the block |
|
|
1
|
|
|
|
|
1
|
|
|
|
1
|
|
|
|
|
703
|
|
|
1300
|
14
|
50
|
|
|
|
7
|
if (!${$_}) { |
|
|
14
|
|
|
|
|
34
|
|
|
1301
|
14
|
|
|
|
|
7
|
${$_} = 0; |
|
|
14
|
|
|
|
|
16
|
|
|
1302
|
|
|
|
|
|
|
} |
|
1303
|
|
|
|
|
|
|
} |
|
1304
|
2
|
50
|
|
|
|
5
|
if (!$multiplet_count) { $multiplet_count = 0; } |
|
|
0
|
|
|
|
|
0
|
|
|
1305
|
2
|
100
|
|
|
|
4
|
if ($total_only) { |
|
1306
|
1
|
|
|
|
|
5
|
return $count; |
|
1307
|
|
|
|
|
|
|
} |
|
1308
|
1
|
|
|
|
|
11
|
$return_string = "Singt/singn/doub/pair/mult/total : ". |
|
1309
|
|
|
|
|
|
|
"$singlets,$singletons,$doublets(". |
|
1310
|
|
|
|
|
|
|
($doublets*2)."),$pairs(".($pairs*2). |
|
1311
|
|
|
|
|
|
|
"),$count_multiplets($multiplet_count),$count"; |
|
1312
|
1
|
|
|
|
|
5
|
return $return_string; |
|
1313
|
|
|
|
|
|
|
} |
|
1314
|
|
|
|
|
|
|
|
|
1315
|
|
|
|
|
|
|
=head2 write_stats() |
|
1316
|
|
|
|
|
|
|
|
|
1317
|
|
|
|
|
|
|
Title : write_stats() |
|
1318
|
|
|
|
|
|
|
Usage : $o_consed->write_stats(); |
|
1319
|
|
|
|
|
|
|
Function: Write a file called "statistics" containing numbers similar to |
|
1320
|
|
|
|
|
|
|
those provided in sum_lets(). |
|
1321
|
|
|
|
|
|
|
Returns : Nothing. Write a file in $o_consed->{path} containing something |
|
1322
|
|
|
|
|
|
|
like this: |
|
1323
|
|
|
|
|
|
|
|
|
1324
|
|
|
|
|
|
|
0,0,50(100),0(0),0(0),100 |
|
1325
|
|
|
|
|
|
|
|
|
1326
|
|
|
|
|
|
|
Where the numbers provided are in the format described in the |
|
1327
|
|
|
|
|
|
|
documentation for sum_lets(). |
|
1328
|
|
|
|
|
|
|
Args : None. |
|
1329
|
|
|
|
|
|
|
Notes : This might break platform independence, I do not know. |
|
1330
|
|
|
|
|
|
|
|
|
1331
|
|
|
|
|
|
|
See L |
|
1332
|
|
|
|
|
|
|
|
|
1333
|
|
|
|
|
|
|
=cut |
|
1334
|
|
|
|
|
|
|
|
|
1335
|
|
|
|
|
|
|
sub write_stats { |
|
1336
|
|
|
|
|
|
|
# worry about platform dependence here? |
|
1337
|
|
|
|
|
|
|
# oh shucksdarn. |
|
1338
|
0
|
|
|
0
|
1
|
0
|
my $self = shift; |
|
1339
|
0
|
|
|
|
|
0
|
my $stats_filename = $self->{'path'}."statistics"; |
|
1340
|
0
|
|
|
|
|
0
|
my $statistics_raw = $self->sum_lets; |
|
1341
|
0
|
|
|
|
|
0
|
my ($statsfilecontents) = $statistics_raw =~ s/.*\ \:\ //g; |
|
1342
|
0
|
|
|
|
|
0
|
umask 0001; |
|
1343
|
0
|
|
|
|
|
0
|
my $fh = Bio::Root::IO->new(-file=>"$stats_filename"); |
|
1344
|
|
|
|
|
|
|
# open my $STATSFILE, '>', $stats_filename or print "Could not write the statsfile: $!\n"); |
|
1345
|
0
|
|
|
|
|
0
|
$fh->_print("$statsfilecontents"); |
|
1346
|
|
|
|
|
|
|
# close $STATSFILE; |
|
1347
|
0
|
|
|
|
|
0
|
$fh->close(); |
|
1348
|
|
|
|
|
|
|
} |
|
1349
|
|
|
|
|
|
|
|
|
1350
|
|
|
|
|
|
|
=head2 get_singletons() |
|
1351
|
|
|
|
|
|
|
|
|
1352
|
|
|
|
|
|
|
Title : get_singletons() |
|
1353
|
|
|
|
|
|
|
Usage : @singletons = $o_consed->get_singletons(); |
|
1354
|
|
|
|
|
|
|
Function: Return the keynames of the singletons. |
|
1355
|
|
|
|
|
|
|
Returns : Returns an array containing the keynames of all |
|
1356
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed sequences in the class "singleton". |
|
1357
|
|
|
|
|
|
|
Args : None. |
|
1358
|
|
|
|
|
|
|
Notes : |
|
1359
|
|
|
|
|
|
|
|
|
1360
|
|
|
|
|
|
|
=cut |
|
1361
|
|
|
|
|
|
|
|
|
1362
|
|
|
|
|
|
|
sub get_singletons { |
|
1363
|
|
|
|
|
|
|
# returns an array of singleton names |
|
1364
|
|
|
|
|
|
|
# singletons are contigs with one member (see consed documentation) |
|
1365
|
3
|
|
|
3
|
1
|
454
|
my $self = shift; |
|
1366
|
3
|
|
|
|
|
3
|
my (@singletons,@array); |
|
1367
|
3
|
|
|
|
|
4
|
foreach my $key (sort keys %{$self->{'contigs'}}) { |
|
|
3
|
|
|
|
|
89
|
|
|
1368
|
354
|
50
|
|
|
|
379
|
if ($self->{'contigs'}->{$key}->{'class'}) { |
|
1369
|
|
|
|
|
|
|
# print ("$key class: $self->{'contigs'}->{$key}->{'class'}\n"); |
|
1370
|
|
|
|
|
|
|
} |
|
1371
|
|
|
|
|
|
|
else { |
|
1372
|
|
|
|
|
|
|
# print("$key belongs to no class. why?\n"); |
|
1373
|
|
|
|
|
|
|
} |
|
1374
|
354
|
50
|
|
|
|
408
|
if ($self->{'contigs'}->{$key}->{'member_array'}) { |
|
1375
|
354
|
|
|
|
|
182
|
@array = @{$self->{'contigs'}->{$key}->{'member_array'}}; |
|
|
354
|
|
|
|
|
494
|
|
|
1376
|
|
|
|
|
|
|
} |
|
1377
|
354
|
|
|
|
|
223
|
my $num_array_elem = @array; |
|
1378
|
354
|
100
|
66
|
|
|
818
|
if ($num_array_elem == 1 && $self->{'contigs'}->{$key}->{'class'} && $self->{'contigs'}->{$key}->{'class'} eq "singleton") { push @singletons,$key; } |
|
|
9
|
|
66
|
|
|
10
|
|
|
1379
|
|
|
|
|
|
|
} |
|
1380
|
3
|
|
|
|
|
18
|
return @singletons; |
|
1381
|
|
|
|
|
|
|
} |
|
1382
|
|
|
|
|
|
|
|
|
1383
|
|
|
|
|
|
|
=head2 get_pairs() |
|
1384
|
|
|
|
|
|
|
|
|
1385
|
|
|
|
|
|
|
Title : get_pairs() |
|
1386
|
|
|
|
|
|
|
Usage : @pairs = $o_consed->get_pairs(); |
|
1387
|
|
|
|
|
|
|
Function: Return the keynames of the pairs. |
|
1388
|
|
|
|
|
|
|
Returns : Returns an array containing the keynames of all |
|
1389
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed sequences in the class "pair". |
|
1390
|
|
|
|
|
|
|
Args : None. |
|
1391
|
|
|
|
|
|
|
Notes : |
|
1392
|
|
|
|
|
|
|
|
|
1393
|
|
|
|
|
|
|
=cut |
|
1394
|
|
|
|
|
|
|
|
|
1395
|
|
|
|
|
|
|
sub get_pairs { |
|
1396
|
|
|
|
|
|
|
# returns an array of pair contig names |
|
1397
|
|
|
|
|
|
|
# a pair is a contig of two where the names do not match |
|
1398
|
3
|
|
|
3
|
1
|
3
|
my $self = shift; |
|
1399
|
3
|
|
|
|
|
3
|
my (@pairs,@array); |
|
1400
|
3
|
|
|
|
|
3
|
foreach my $key (sort keys %{$self->{'contigs'}}) { |
|
|
3
|
|
|
|
|
95
|
|
|
1401
|
354
|
50
|
|
|
|
418
|
if ($self->{'contigs'}->{$key}->{'member_array'}) { |
|
1402
|
354
|
100
|
100
|
|
|
197
|
if (@{$self->{'contigs'}->{$key}->{'member_array'}} == 2 && |
|
|
354
|
|
|
|
|
705
|
|
|
1403
|
|
|
|
|
|
|
$self->{'contigs'}->{$key}->{'class'} eq "pair") { |
|
1404
|
3
|
|
|
|
|
7
|
push @pairs,$key; |
|
1405
|
|
|
|
|
|
|
} |
|
1406
|
|
|
|
|
|
|
} |
|
1407
|
|
|
|
|
|
|
} |
|
1408
|
3
|
|
|
|
|
15
|
return @pairs; |
|
1409
|
|
|
|
|
|
|
} |
|
1410
|
|
|
|
|
|
|
|
|
1411
|
|
|
|
|
|
|
=head2 get_name($contig_keyname) |
|
1412
|
|
|
|
|
|
|
|
|
1413
|
|
|
|
|
|
|
Title : get_name($contig_keyname) |
|
1414
|
|
|
|
|
|
|
Usage : $name = $o_consed->get_name($contig_keyname); |
|
1415
|
|
|
|
|
|
|
Function: Return the {name} for $contig_keyname. |
|
1416
|
|
|
|
|
|
|
Returns : A string. ({name}) |
|
1417
|
|
|
|
|
|
|
Args : A contig keyname. |
|
1418
|
|
|
|
|
|
|
Notes : |
|
1419
|
|
|
|
|
|
|
|
|
1420
|
|
|
|
|
|
|
=cut |
|
1421
|
|
|
|
|
|
|
|
|
1422
|
|
|
|
|
|
|
sub get_name { |
|
1423
|
0
|
|
|
0
|
1
|
0
|
my ($self,$contig) = @_; |
|
1424
|
0
|
|
|
|
|
0
|
return $self->{'contigs'}->{$contig}->{'name'}; |
|
1425
|
|
|
|
|
|
|
} |
|
1426
|
|
|
|
|
|
|
|
|
1427
|
|
|
|
|
|
|
=head2 _get_contig_name(\@array_containing_reads) |
|
1428
|
|
|
|
|
|
|
|
|
1429
|
|
|
|
|
|
|
Title : _get_contig_name(\@array_containing_reads) |
|
1430
|
|
|
|
|
|
|
Usage : $o_consed->_get_contig_name(\@array_containing_reads); |
|
1431
|
|
|
|
|
|
|
Function: The logic for the set_doublets subroutine. |
|
1432
|
|
|
|
|
|
|
Returns : The name for this contig. |
|
1433
|
|
|
|
|
|
|
Args : A reference to an array containing read names. |
|
1434
|
|
|
|
|
|
|
Notes : Depends on reverse_designator. Be sure this is set the way you |
|
1435
|
|
|
|
|
|
|
intend. |
|
1436
|
|
|
|
|
|
|
|
|
1437
|
|
|
|
|
|
|
=cut |
|
1438
|
|
|
|
|
|
|
|
|
1439
|
|
|
|
|
|
|
sub _get_contig_name { |
|
1440
|
46
|
|
|
46
|
|
32
|
my ($self,$r_array) = @_; |
|
1441
|
46
|
|
|
|
|
73
|
my @contig_members = @$r_array; |
|
1442
|
46
|
|
|
|
|
25
|
my @name_nodir; |
|
1443
|
46
|
|
|
|
|
44
|
foreach (@contig_members) { |
|
1444
|
|
|
|
|
|
|
# how can I distinguish the clone name from the direction label? |
|
1445
|
|
|
|
|
|
|
# look for $Consed::reverse_designator and $Consed::forward_designator |
|
1446
|
|
|
|
|
|
|
# what if you do not find _any_ of those? |
|
1447
|
92
|
|
50
|
|
|
114
|
my $forward_designator = $self->{'forward_designator'} || "f"; |
|
1448
|
92
|
|
50
|
|
|
101
|
my $reverse_designator = $self->{'reverse_designator'} || "r"; |
|
1449
|
92
|
|
33
|
|
|
406
|
my $any_hits = /(.+)($forward_designator.*)/ || /(.+)($reverse_designator.*)/||/(.+)(_.+)/; |
|
1450
|
92
|
|
|
|
|
97
|
my $name = $1; |
|
1451
|
92
|
|
|
|
|
65
|
my $suffix = $2; |
|
1452
|
92
|
50
|
|
|
|
110
|
if ($name) { |
|
1453
|
|
|
|
|
|
|
# print("\t\$name is $name "); |
|
1454
|
|
|
|
|
|
|
} |
|
1455
|
92
|
50
|
|
|
|
94
|
if ($suffix) { |
|
1456
|
|
|
|
|
|
|
# print("and \$suffix is $suffix.\n"); |
|
1457
|
|
|
|
|
|
|
} |
|
1458
|
|
|
|
|
|
|
# Jee, I hope we get a naming convention soon |
|
1459
|
92
|
50
|
|
|
|
106
|
if ($suffix) { |
|
1460
|
92
|
50
|
66
|
|
|
314
|
if ($suffix =~ /^$forward_designator/ || $suffix =~ /^$reverse_designator/) { |
|
1461
|
92
|
|
|
|
|
136
|
push @name_nodir,$name; |
|
1462
|
|
|
|
|
|
|
} |
|
1463
|
|
|
|
|
|
|
# bugwatch here! should this be unnested? |
|
1464
|
|
|
|
|
|
|
else { |
|
1465
|
0
|
|
|
|
|
0
|
push @name_nodir,"$name$suffix"; |
|
1466
|
|
|
|
|
|
|
} |
|
1467
|
|
|
|
|
|
|
} |
|
1468
|
|
|
|
|
|
|
} |
|
1469
|
|
|
|
|
|
|
# print("\@name_nodir: @name_nodir\n"); |
|
1470
|
46
|
|
|
|
|
42
|
my $mismatch = 0; |
|
1471
|
46
|
|
|
|
|
67
|
for (my $counter=0; $counter<@name_nodir;$counter++) { |
|
1472
|
92
|
100
|
|
|
|
179
|
next if ($name_nodir[0] eq $name_nodir[$counter]); |
|
1473
|
1
|
|
|
|
|
3
|
$mismatch = 1; |
|
1474
|
|
|
|
|
|
|
} |
|
1475
|
46
|
100
|
|
|
|
55
|
if ($mismatch == 0) { |
|
1476
|
|
|
|
|
|
|
# print("\tYou have a cohesive contig named $name_nodir[0].\n\n"); |
|
1477
|
45
|
|
|
|
|
64
|
return $name_nodir[0]; |
|
1478
|
|
|
|
|
|
|
} else { |
|
1479
|
|
|
|
|
|
|
# print("\tYou have mixed names in this contig.\n\n"); |
|
1480
|
|
|
|
|
|
|
} |
|
1481
|
|
|
|
|
|
|
} # end _get_contig_name |
|
1482
|
|
|
|
|
|
|
|
|
1483
|
|
|
|
|
|
|
=head2 get_doublets() |
|
1484
|
|
|
|
|
|
|
|
|
1485
|
|
|
|
|
|
|
Title : get_doublets() |
|
1486
|
|
|
|
|
|
|
Usage : @doublets = $o_consed->get_doublets(); |
|
1487
|
|
|
|
|
|
|
Function: Return the keynames of the doublets. |
|
1488
|
|
|
|
|
|
|
Returns : Returns an array containing the keynames of all |
|
1489
|
|
|
|
|
|
|
Bio::Tools::Alignment::Consed sequences in the class "doublet". |
|
1490
|
|
|
|
|
|
|
Args : None. |
|
1491
|
|
|
|
|
|
|
Notes : |
|
1492
|
|
|
|
|
|
|
|
|
1493
|
|
|
|
|
|
|
=cut |
|
1494
|
|
|
|
|
|
|
|
|
1495
|
|
|
|
|
|
|
sub get_doublets { |
|
1496
|
3
|
|
|
3
|
1
|
5
|
my $self = shift; |
|
1497
|
3
|
50
|
|
|
|
9
|
if (!$self->{doublets_set}) { |
|
1498
|
0
|
|
|
|
|
0
|
$self->warn("You need to set the doublets before you can get them. Doing that now."); |
|
1499
|
0
|
|
|
|
|
0
|
$self->set_doublets(); |
|
1500
|
|
|
|
|
|
|
} |
|
1501
|
3
|
|
|
|
|
1
|
my @doublets; |
|
1502
|
3
|
|
|
|
|
5
|
foreach (sort keys %{$self->{'contigs'}}) { |
|
|
3
|
|
|
|
|
96
|
|
|
1503
|
354
|
100
|
100
|
|
|
927
|
if ($self->{'contigs'}->{$_}->{name} && $self->{'contigs'}->{$_}->{'class'} eq "doublet") { |
|
1504
|
135
|
|
|
|
|
99
|
push @doublets,$_; |
|
1505
|
|
|
|
|
|
|
} |
|
1506
|
|
|
|
|
|
|
} |
|
1507
|
3
|
|
|
|
|
27
|
return @doublets; |
|
1508
|
|
|
|
|
|
|
} # end get_doublets |
|
1509
|
|
|
|
|
|
|
|
|
1510
|
|
|
|
|
|
|
=head2 dump_hash() |
|
1511
|
|
|
|
|
|
|
|
|
1512
|
|
|
|
|
|
|
Title : dump_hash() |
|
1513
|
|
|
|
|
|
|
Usage : $o_consed->dump_hash(); |
|
1514
|
|
|
|
|
|
|
Function: Use dumpvar.pl to dump out the Bio::Tools::Alignment::Consed |
|
1515
|
|
|
|
|
|
|
object to STDOUT. |
|
1516
|
|
|
|
|
|
|
Returns : Nothing. |
|
1517
|
|
|
|
|
|
|
Args : None. |
|
1518
|
|
|
|
|
|
|
Notes : I used this a lot in debugging. |
|
1519
|
|
|
|
|
|
|
|
|
1520
|
|
|
|
|
|
|
=cut |
|
1521
|
|
|
|
|
|
|
|
|
1522
|
|
|
|
|
|
|
sub dump_hash { |
|
1523
|
0
|
|
|
0
|
1
|
|
my $self = shift; |
|
1524
|
0
|
|
|
|
|
|
my $dumper = Dumpvalue->new(); |
|
1525
|
0
|
|
|
|
|
|
$self->debug( "Bio::Tools::Alignment::Consed::dump_hash - ". |
|
1526
|
|
|
|
|
|
|
"The following is the contents of the contig hash...\n"); |
|
1527
|
0
|
|
|
|
|
|
$dumper->dumpValue($self->{'contigs'}); |
|
1528
|
|
|
|
|
|
|
} |
|
1529
|
|
|
|
|
|
|
|
|
1530
|
|
|
|
|
|
|
=head2 dump_hash_compact() |
|
1531
|
|
|
|
|
|
|
|
|
1532
|
|
|
|
|
|
|
Title : dump_hash_compact() |
|
1533
|
|
|
|
|
|
|
Usage : $o_consed->dump_hash_compact(); |
|
1534
|
|
|
|
|
|
|
Function: Dump out the Bio::Tools::Alignment::Consed object in a compact way. |
|
1535
|
|
|
|
|
|
|
Returns : Nothing. |
|
1536
|
|
|
|
|
|
|
Args : Nothing. |
|
1537
|
|
|
|
|
|
|
Notes : Cleaner then dumpValue(), dumpHash(). I used this a lot in |
|
1538
|
|
|
|
|
|
|
debugging. |
|
1539
|
|
|
|
|
|
|
|
|
1540
|
|
|
|
|
|
|
=cut |
|
1541
|
|
|
|
|
|
|
|
|
1542
|
|
|
|
|
|
|
sub dump_hash_compact { |
|
1543
|
1
|
|
|
1
|
|
5
|
no strict 'refs'; # renege for the block |
|
|
1
|
|
|
|
|
0
|
|
|
|
1
|
|
|
|
|
1080
|
|
|
1544
|
0
|
|
|
0
|
1
|
|
my ($self,$sequence) = @_; |
|
1545
|
|
|
|
|
|
|
# get the classes |
|
1546
|
0
|
|
|
|
|
|
my @singlets = $self->get_singlets(); |
|
1547
|
0
|
|
|
|
|
|
my @singletons = $self->get_singletons(); |
|
1548
|
0
|
|
|
|
|
|
my @doublets = $self->get_doublets(); |
|
1549
|
0
|
|
|
|
|
|
my @pairs = $self->get_pairs(); |
|
1550
|
0
|
|
|
|
|
|
my @multiplets = $self->get_multiplets(); |
|
1551
|
0
|
|
|
|
|
|
print("Name\tClass\tMembers\tQuality?\n"); |
|
1552
|
0
|
|
|
|
|
|
foreach (@singlets) { |
|
1553
|
0
|
|
|
|
|
|
my @members = $self->get_members($_); |
|
1554
|
0
|
|
|
|
|
|
print($self->get_name($_)."\tsinglets\t".(join',',@members)."\t"); |
|
1555
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$_}->{'quality'}) { |
|
1556
|
0
|
|
|
|
|
|
print("qualities found here\n"); |
|
1557
|
|
|
|
|
|
|
} else { |
|
1558
|
0
|
|
|
|
|
|
print("no qualities found here\n"); |
|
1559
|
|
|
|
|
|
|
} |
|
1560
|
|
|
|
|
|
|
|
|
1561
|
|
|
|
|
|
|
} |
|
1562
|
0
|
|
|
|
|
|
foreach (@singletons) { |
|
1563
|
0
|
|
|
|
|
|
my @members = $self->get_members($_); |
|
1564
|
0
|
|
|
|
|
|
print($self->get_name($_)."\tsingletons\t".(join',',@members)."\t"); |
|
1565
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$_}->{'quality'}) { |
|
1566
|
0
|
|
|
|
|
|
print("qualities found here\n"); |
|
1567
|
|
|
|
|
|
|
} else { |
|
1568
|
0
|
|
|
|
|
|
print("no qualities found here\n"); |
|
1569
|
|
|
|
|
|
|
} |
|
1570
|
|
|
|
|
|
|
} |
|
1571
|
0
|
|
|
|
|
|
foreach my $pair (@pairs) { |
|
1572
|
0
|
|
|
|
|
|
my @members = $self->get_members($pair); |
|
1573
|
0
|
|
|
|
|
|
my $name; |
|
1574
|
0
|
0
|
|
|
|
|
if (!$self->get_name($pair)) { |
|
1575
|
0
|
|
|
|
|
|
$name = "BLANK"; |
|
1576
|
|
|
|
|
|
|
} else { |
|
1577
|
0
|
|
|
|
|
|
$name = $self->get_name($pair); |
|
1578
|
|
|
|
|
|
|
} |
|
1579
|
0
|
|
|
|
|
|
print("$name\tpairs\t".(join',',@members)."\n"); |
|
1580
|
|
|
|
|
|
|
} |
|
1581
|
0
|
|
|
|
|
|
foreach (@doublets) { |
|
1582
|
0
|
|
|
|
|
|
my @members = $self->get_members_by_name($_); |
|
1583
|
0
|
|
|
|
|
|
print("$_\tdoublets\t".(join',',@members)."\t"); |
|
1584
|
0
|
|
|
|
|
|
my $contig_number = &get_contig_number_by_name($self,$_); |
|
1585
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$contig_number}->{'quality'}) { |
|
1586
|
0
|
|
|
|
|
|
print("qualities found here\n"); |
|
1587
|
|
|
|
|
|
|
} else { |
|
1588
|
0
|
|
|
|
|
|
print("no qualities found here\n"); |
|
1589
|
|
|
|
|
|
|
} |
|
1590
|
|
|
|
|
|
|
# print($_."\tdoublets\t".(join',',@members)."\n"); |
|
1591
|
|
|
|
|
|
|
} |
|
1592
|
0
|
|
|
|
|
|
foreach (@multiplets) { |
|
1593
|
0
|
|
|
|
|
|
my @members = $self->get_members($_); |
|
1594
|
0
|
|
|
|
|
|
print("Contig $_"."\tmultiplets\t".(join',',@members)."\n"); |
|
1595
|
|
|
|
|
|
|
} |
|
1596
|
|
|
|
|
|
|
} # end dump_hash_compact |
|
1597
|
|
|
|
|
|
|
|
|
1598
|
|
|
|
|
|
|
=head2 get_phreds() |
|
1599
|
|
|
|
|
|
|
|
|
1600
|
|
|
|
|
|
|
Title : get_phreds() |
|
1601
|
|
|
|
|
|
|
Usage : @phreds = $o_consed->get_phreds(); |
|
1602
|
|
|
|
|
|
|
Function: For each doublet in the Bio::Tools::Alignment::Consed hash, go |
|
1603
|
|
|
|
|
|
|
and get the phreds for the top and bottom reads. Place them into |
|
1604
|
|
|
|
|
|
|
{top_phreds} and {bottom_phreds}. |
|
1605
|
|
|
|
|
|
|
Returns : Nothing. |
|
1606
|
|
|
|
|
|
|
Args : Nothing. |
|
1607
|
|
|
|
|
|
|
|
|
1608
|
|
|
|
|
|
|
Requires parse_phd() and reverse_and_complement(). I realize that it |
|
1609
|
|
|
|
|
|
|
would be much more elegant to pull qualities as required but there |
|
1610
|
|
|
|
|
|
|
were certain "features" in the acefile that required a bit more |
|
1611
|
|
|
|
|
|
|
detailed work be done to get the qualities for certain parts of the |
|
1612
|
|
|
|
|
|
|
consensus sequence. In order to make _sure_ that this was done |
|
1613
|
|
|
|
|
|
|
properly I wrote things to do all steps and then I used dump_hash() |
|
1614
|
|
|
|
|
|
|
and checked each one to ensure expected behavior. I have never changed |
|
1615
|
|
|
|
|
|
|
this, so there you are. |
|
1616
|
|
|
|
|
|
|
|
|
1617
|
|
|
|
|
|
|
=cut |
|
1618
|
|
|
|
|
|
|
|
|
1619
|
|
|
|
|
|
|
sub get_phreds { |
|
1620
|
|
|
|
|
|
|
# this subroutine is the target of a rewrite to use the Bio::Tools::Alignment::Phred object. |
|
1621
|
0
|
|
|
0
|
1
|
|
my $self = shift; |
|
1622
|
0
|
|
|
|
|
|
my $current_contig; |
|
1623
|
0
|
|
|
|
|
|
foreach $current_contig (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
|
|
|
1624
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$current_contig}->{'class'} eq "doublet") { |
|
1625
|
0
|
|
|
|
|
|
$self->debug("$current_contig is a doublet. Going to parse_phd for top($self->{'contigs'}->{$current_contig}->{'top_name'}) and bottom($self->{'contigs'}->{$current_contig}->{'bottom_name'})\n"); |
|
1626
|
0
|
|
|
|
|
|
my $r_phreds_top = &parse_phd($self,$self->{'contigs'}->{$current_contig}->{'top_name'}); |
|
1627
|
0
|
|
|
|
|
|
my $r_phreds_bottom = &parse_phd($self,$self->{'contigs'}->{$current_contig}->{'bottom_name'}); |
|
1628
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$current_contig}->{'top_complement'} eq "C") { |
|
1629
|
|
|
|
|
|
|
# print("Reversing and complementing...\n"); |
|
1630
|
0
|
|
|
|
|
|
$r_phreds_top = &reverse_and_complement($r_phreds_top); |
|
1631
|
|
|
|
|
|
|
} |
|
1632
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$current_contig}->{'bottom_complement'} eq "C") { |
|
1633
|
0
|
|
|
|
|
|
$r_phreds_bottom = &reverse_and_complement($r_phreds_bottom); |
|
1634
|
|
|
|
|
|
|
} |
|
1635
|
0
|
|
|
|
|
|
$self->{'contigs'}->{$current_contig}->{'top_phreds'} = $r_phreds_top; |
|
1636
|
0
|
|
|
|
|
|
$self->{'contigs'}->{$current_contig}->{'bottom_phreds'} = $r_phreds_bottom; |
|
1637
|
|
|
|
|
|
|
} |
|
1638
|
|
|
|
|
|
|
} |
|
1639
|
|
|
|
|
|
|
} |
|
1640
|
|
|
|
|
|
|
|
|
1641
|
|
|
|
|
|
|
=head2 parse_phd($read_name) |
|
1642
|
|
|
|
|
|
|
|
|
1643
|
|
|
|
|
|
|
Title : parse_phd($read_name) |
|
1644
|
|
|
|
|
|
|
Usage : $o_consed->parse_phd($read_name); |
|
1645
|
|
|
|
|
|
|
Function: Suck in the contents of a .phd file. |
|
1646
|
|
|
|
|
|
|
Returns : A reference to an array containing the quality values for the read. |
|
1647
|
|
|
|
|
|
|
Args : The name of a read. |
|
1648
|
|
|
|
|
|
|
Notes : This is a significantly weak subroutine because it was always |
|
1649
|
|
|
|
|
|
|
intended that these functions, along with the functions provided by |
|
1650
|
|
|
|
|
|
|
get_phreds() be put into the Bio::SeqIO:phd module. This is done |
|
1651
|
|
|
|
|
|
|
now but the Bio::Tools::Alignment::Consed module has not be |
|
1652
|
|
|
|
|
|
|
rewritten to reflect this change. |
|
1653
|
|
|
|
|
|
|
|
|
1654
|
|
|
|
|
|
|
See L for more information. |
|
1655
|
|
|
|
|
|
|
|
|
1656
|
|
|
|
|
|
|
=cut |
|
1657
|
|
|
|
|
|
|
|
|
1658
|
|
|
|
|
|
|
sub parse_phd { |
|
1659
|
0
|
|
|
0
|
1
|
|
my ($self,$sequence_name) = @_; |
|
1660
|
0
|
|
|
|
|
|
$self->debug("Parsing phd for $sequence_name\n"); |
|
1661
|
0
|
|
|
|
|
|
my $in_dna = 0; |
|
1662
|
0
|
|
|
|
|
|
my $base_number = 0; |
|
1663
|
0
|
|
|
|
|
|
my (@bases,@current_line); |
|
1664
|
|
|
|
|
|
|
# print("parse_phd: $sequence_name\n"); |
|
1665
|
0
|
|
|
|
|
|
my $fh = Bio::Root::IO->new |
|
1666
|
|
|
|
|
|
|
(-file=>"$self->{path}/../phd_dir/$sequence_name.phd.1"); |
|
1667
|
0
|
|
|
|
|
|
while ($fh->_readline()) { |
|
1668
|
|
|
|
|
|
|
# print("Reading a line from a phredfile!\n"); |
|
1669
|
0
|
|
|
|
|
|
chomp; |
|
1670
|
0
|
0
|
|
|
|
|
if (/^BEGIN_DNA/) { $in_dna = 1; next} |
|
|
0
|
|
|
|
|
|
|
|
|
0
|
|
|
|
|
|
|
|
1671
|
0
|
0
|
|
|
|
|
if (/^END_DNA/) { last; } |
|
|
0
|
|
|
|
|
|
|
|
1672
|
0
|
0
|
|
|
|
|
if (!$in_dna) { next; } |
|
|
0
|
|
|
|
|
|
|
|
1673
|
0
|
|
|
|
|
|
push(@bases,$_); |
|
1674
|
|
|
|
|
|
|
} |
|
1675
|
0
|
|
|
|
|
|
return \@bases; |
|
1676
|
|
|
|
|
|
|
} |
|
1677
|
|
|
|
|
|
|
|
|
1678
|
|
|
|
|
|
|
=head2 reverse_and_complement(\@source) |
|
1679
|
|
|
|
|
|
|
|
|
1680
|
|
|
|
|
|
|
Title : reverse_and_complement(\@source) |
|
1681
|
|
|
|
|
|
|
Usage : $reference_to_array = $o_consed->reverse_and_complement(\@source); |
|
1682
|
|
|
|
|
|
|
Function: A stub for the recursive routine reverse_recurse(). |
|
1683
|
|
|
|
|
|
|
Returns : A reference to a reversed and complemented array of phred data. |
|
1684
|
|
|
|
|
|
|
Args : A reference to an array of phred data. |
|
1685
|
|
|
|
|
|
|
Notes : |
|
1686
|
|
|
|
|
|
|
|
|
1687
|
|
|
|
|
|
|
=cut |
|
1688
|
|
|
|
|
|
|
|
|
1689
|
|
|
|
|
|
|
sub reverse_and_complement { |
|
1690
|
0
|
|
|
0
|
1
|
|
my $r_source = shift; |
|
1691
|
0
|
|
|
|
|
|
my $r_destination; |
|
1692
|
0
|
|
|
|
|
|
$r_destination = &reverse_recurse($r_source,$r_destination); |
|
1693
|
0
|
|
|
|
|
|
return $r_destination; |
|
1694
|
|
|
|
|
|
|
} |
|
1695
|
|
|
|
|
|
|
|
|
1696
|
|
|
|
|
|
|
=head2 reverse_recurse($r_source,$r_destination) |
|
1697
|
|
|
|
|
|
|
|
|
1698
|
|
|
|
|
|
|
Title : reverse_recurse(\@source,\@destination) |
|
1699
|
|
|
|
|
|
|
Usage : $o_consed->reverse_recurse(\@source,\@destination); |
|
1700
|
|
|
|
|
|
|
Function: A recursive routine to reverse and complement an array of |
|
1701
|
|
|
|
|
|
|
phred data. |
|
1702
|
|
|
|
|
|
|
Returns : A reference to an array containing reversed phred data. |
|
1703
|
|
|
|
|
|
|
Args : A reference to a source array and a reverence to a destination |
|
1704
|
|
|
|
|
|
|
array. |
|
1705
|
|
|
|
|
|
|
|
|
1706
|
|
|
|
|
|
|
Recursion is kewl, but this sub should likely be _reverse_recurse. |
|
1707
|
|
|
|
|
|
|
|
|
1708
|
|
|
|
|
|
|
=cut |
|
1709
|
|
|
|
|
|
|
|
|
1710
|
|
|
|
|
|
|
|
|
1711
|
|
|
|
|
|
|
sub reverse_recurse($$) { |
|
1712
|
0
|
|
|
0
|
1
|
|
my ($r_source,$r_destination) = @_; |
|
1713
|
0
|
0
|
|
|
|
|
if (!@$r_source) { |
|
1714
|
0
|
|
|
|
|
|
return $r_destination; |
|
1715
|
|
|
|
|
|
|
} |
|
1716
|
0
|
|
|
|
|
|
$_=pop(@$r_source); |
|
1717
|
0
|
0
|
0
|
|
|
|
s/c/g/ || s/g/c/ || s/a/t/ || s/t/a/; |
|
|
|
|
0
|
|
|
|
|
|
1718
|
0
|
|
|
|
|
|
push(@$r_destination,$_); |
|
1719
|
0
|
|
|
|
|
|
&reverse_recurse($r_source,$r_destination); |
|
1720
|
|
|
|
|
|
|
} |
|
1721
|
|
|
|
|
|
|
|
|
1722
|
|
|
|
|
|
|
=head2 show_missing_sequence() |
|
1723
|
|
|
|
|
|
|
|
|
1724
|
|
|
|
|
|
|
Title : show_missing_sequence(); |
|
1725
|
|
|
|
|
|
|
Usage : $o_consed->show_missing_sequence(); |
|
1726
|
|
|
|
|
|
|
Function: Used by set_trim_points_doublets() to fill in quality values where |
|
1727
|
|
|
|
|
|
|
consed (phrap?) set them to 0 at the beginning and/or end of the |
|
1728
|
|
|
|
|
|
|
consensus sequences. |
|
1729
|
|
|
|
|
|
|
Returns : Nothing. |
|
1730
|
|
|
|
|
|
|
Args : None. |
|
1731
|
|
|
|
|
|
|
|
|
1732
|
|
|
|
|
|
|
Acts on doublets only. Really very somewhat quite ugly. A disgusting |
|
1733
|
|
|
|
|
|
|
kludge. I It was written stepwise with no real plan |
|
1734
|
|
|
|
|
|
|
because it was not really evident why consed (phrap?) was doing this. |
|
1735
|
|
|
|
|
|
|
|
|
1736
|
|
|
|
|
|
|
=cut |
|
1737
|
|
|
|
|
|
|
|
|
1738
|
|
|
|
|
|
|
sub show_missing_sequence() { |
|
1739
|
|
|
|
|
|
|
|
|
1740
|
|
|
|
|
|
|
# decide which sequence should not have been clipped at consensus |
|
1741
|
|
|
|
|
|
|
# position = 0 |
|
1742
|
|
|
|
|
|
|
|
|
1743
|
0
|
|
|
0
|
1
|
|
my $self = shift; |
|
1744
|
0
|
|
|
|
|
|
&get_phreds($self); |
|
1745
|
0
|
|
|
|
|
|
my ($current_contig,@qualities); |
|
1746
|
0
|
|
|
|
|
|
foreach $current_contig (sort keys %{$self->{'contigs'}}) { |
|
|
0
|
|
|
|
|
|
|
|
1747
|
0
|
0
|
|
|
|
|
if ($self->{'contigs'}->{$current_contig}->{'class'} eq "doublet") { |
|
1748
|
0
|
|
|
|
|
|
my $number_leading_xs = 0; |
|
1749
|
0
|
|
|
|
|
|
my $number_trailing_xs = 0; |
|
1750
|
0
|
|
|
|
|
|
my $measurer = $self->{'contigs'}->{$current_contig}->{'quality'}; |
|
1751
|
0
|
|
|
|
|
|
while ($measurer =~ s/^\ 0\ /\ /) { |
|
1752
|
0
|
|
|
|
|
|
$number_leading_xs++; |
|
1753
|
|
|
|
|
|
|
} |
|
1754
|
0
|
|
|
|
|
|
while ($measurer =~ s/\ 0(\s*)$/$1/) { |
|
1755
|
0
|
|
|
|
|
|
$number_trailing_xs++; |
|
1756
|
|
|
|
|
|
|
} |
|
1757
|
0
|
|
|
|
|
|
@qualities = split(' ',$self->{'contigs'}->{$current_contig}->{'quality'}); |
|
1758
|
0
|
|
|
|
|
|
my $in_initial_zeros = 0; |
|
1759
|
0
|
|
|
|
|
|
for (my $count=0;$count
|
|
1760
|
0
|
0
|
|
|
|
|
if ($qualities[$count] == 0) { |
|
1761
|
0
|
|
|
|
|
|
my ($quality,$top_phred_position,$bottom_phred_position,$top_phred_data,$bottom_phred_data); |
|
1762
|
|
|
|
|
|
|
# print("The quality of the consensus at ".($count+1)." is zero. Retrieving the real quality value.\n"); |
|
1763
|
|
|
|
|
|
|
# how do I know which strand to get these quality values from???? |
|
1764
|
|
|
|
|
|
|
# boggle |
|
1765
|
0
|
|
|
|
|
|
my $top_quality_here = $self->{'contigs'}->{$current_contig}->{'top_phreds'}->[0-$self->{'contigs'}->{$current_contig}->{'top_start'}+$count+1]; |
|
1766
|
0
|
|
|
|
|
|
my $bottom_quality_here = $self->{'contigs'}->{$current_contig}->{'bottom_phreds'}->[1-$self->{'contigs'}->{$current_contig}->{'bottom_start'}+$count]; |
|
1767
|
0
|
0
|
0
|
|
|
|
if (!$bottom_quality_here || (1-$self->{'contigs'}->{$current_contig}->{'bottom_start'}+$count)<0) { |
|
1768
|
0
|
|
|
|
|
|
$bottom_quality_here = "not found"; |
|
1769
|
|
|
|
|
|
|
} |
|
1770
|
0
|
0
|
|
|
|
|
if (!$top_quality_here) { |
|
1771
|
0
|
|
|
|
|
|
$top_quality_here = "not found"; |
|
1772
|
|
|
|
|
|
|
} |
|
1773
|
|
|
|
|
|
|
# print("Looking for quals at position $count of $current_contig: top position ".(0-$self->{'contigs'}->{$current_contig}->{top_start}+$count)." ($self->{'contigs'}->{$current_contig}->{top_name}) $top_quality_here , bottom position ".(1-$self->{'contigs'}->{$current_contig}->{bottom_start}+$count)." ($self->{'contigs'}->{$current_contig}->{bottom_name}) $bottom_quality_here\n"); |
|
1774
|
0
|
0
|
|
|
|
|
if ($count<$number_leading_xs) { |
|
1775
|
|
|
|
|
|
|
# print("$count is less then $number_leading_xs so I will get the quality from the top strand\n"); |
|
1776
|
|
|
|
|
|
|
# print("retrieved quality is ".$self->{'contigs'}->{$current_contig}->{top_phreds}[0-$self->{'contigs'}->{$current_contig}->{top_start}+$count+1]."\n"); |
|
1777
|
0
|
|
|
|
|
|
my $quality = $top_quality_here; |
|
1778
|
0
|
|
|
|
|
|
$quality =~ /\S+\s(\d+)\s+/; |
|
1779
|
0
|
|
|
|
|
|
$quality = $1; |
|
1780
|
|
|
|
|
|
|
# print("retrieved quality for leading zero $count is $quality\n"); |
|
1781
|
|
|
|
|
|
|
# t 9 9226 |
|
1782
|
0
|
|
|
|
|
|
$qualities[$count] = $quality; |
|
1783
|
|
|
|
|
|
|
} else { |
|
1784
|
|
|
|
|
|
|
# this part is tricky |
|
1785
|
|
|
|
|
|
|
# if the contig is like this |
|
1786
|
|
|
|
|
|
|
# cccccccccccccccc |
|
1787
|
|
|
|
|
|
|
# ffffffffffffffffff |
|
1788
|
|
|
|
|
|
|
# rrrrrrrrrrrrrrrrr |
|
1789
|
|
|
|
|
|
|
# then take the quality value for the trailing zeros in the cons. seq from the r |
|
1790
|
|
|
|
|
|
|
# |
|
1791
|
|
|
|
|
|
|
# but if the contig is like this |
|
1792
|
|
|
|
|
|
|
# cccccccccccccccccc |
|
1793
|
|
|
|
|
|
|
# ffffffffffffffffffffffffffffffff |
|
1794
|
|
|
|
|
|
|
# rrrrrrrrrrrrrrrrrrrrrrrxxxxxxxxr |
|
1795
|
|
|
|
|
|
|
# ^^^ |
|
1796
|
|
|
|
|
|
|
# then any zeros that fall in the positions (^) must be decided whether the quality |
|
1797
|
|
|
|
|
|
|
# is the qual from the f or r strand. I will use the greater number |
|
1798
|
|
|
|
|
|
|
# does a similar situation exist for the leading zeros? i dunno |
|
1799
|
|
|
|
|
|
|
# |
|
1800
|
|
|
|
|
|
|
# print("$count is greater then $number_leading_xs so I will get the quality from the bottom strand\n"); |
|
1801
|
|
|
|
|
|
|
# print("retrieved quality is ".$contigs->{$current_contig}->{top_phreds}[0-$contigs->{$current_contig}->{top_start}+$count+1]."\n"); |
|
1802
|
|
|
|
|
|
|
# my ($quality,$top_phred_position,$bottom_phred_position,$top_phred_data,$bottom_phred_data); |
|
1803
|
0
|
0
|
|
|
|
|
if ($bottom_quality_here eq "not found") { |
|
|
|
0
|
|
|
|
|
|
|
1804
|
|
|
|
|
|
|
# $top_phred_position = 1-$contigs->{$current_contig}->{bottom_start}+$count; |
|
1805
|
|
|
|
|
|
|
# print("Going to get quality from here: $top_phred_position of the top.\n"); |
|
1806
|
|
|
|
|
|
|
# my $temp_quality - $contigs->{$current_contig}->{top_phreds} |
|
1807
|
|
|
|
|
|
|
# $quality = $contigs->{$current_contig}->{top_phreds}[$top_phred_position]; |
|
1808
|
0
|
|
|
|
|
|
$top_quality_here =~ /\w+\s(\d+)\s/; |
|
1809
|
0
|
|
|
|
|
|
$quality = $1; |
|
1810
|
|
|
|
|
|
|
} elsif ($top_quality_here eq "not found") { |
|
1811
|
|
|
|
|
|
|
# $bottom_phred_position = 1+$contigs->{$current_contig}->{bottom_start}+$count; |
|
1812
|
|
|
|
|
|
|
# print("Going to get quality from here: $bottom_phred_position of the bottom.\n"); |
|
1813
|
|
|
|
|
|
|
# $quality = $contigs->{$current_contig}->{bottom_phreds}[$bottom_phred_position]; |
|
1814
|
|
|
|
|
|
|
# print("Additional: no top quality but bottom is $quality\n"); |
|
1815
|
0
|
|
|
|
|
|
$bottom_quality_here =~ /\w+\s(\d+)\s/; |
|
1816
|
0
|
|
|
|
|
|
$quality = $1; |
|
1817
|
|
|
|
|
|
|
} else { |
|
1818
|
|
|
|
|
|
|
# print("Oh jeepers, there are 2 qualities to choose from at this position.\n"); |
|
1819
|
|
|
|
|
|
|
# print("Going to compare these phred qualities: top: #$top_quality_here# bottom: #$bottom_quality_here#\n"); |
|
1820
|
|
|
|
|
|
|
# now you have to compare them |
|
1821
|
|
|
|
|
|
|
# my $top_quality_phred = $contigs->{$current_contig}->{top_phreds}[$top_phred_position]; |
|
1822
|
|
|
|
|
|
|
# #t 40 875# |
|
1823
|
|
|
|
|
|
|
# print("regexing #$top_quality_here#... "); |
|
1824
|
0
|
|
|
|
|
|
$top_quality_here =~ /\w\ (\d+)\s/; |
|
1825
|
0
|
|
|
|
|
|
my $top_quality = $1; |
|
1826
|
|
|
|
|
|
|
# print("$top_quality\nregexing #$bottom_quality_here#... "); |
|
1827
|
0
|
|
|
|
|
|
$bottom_quality_here =~ /\w\ (\d+)\s/; |
|
1828
|
0
|
|
|
|
|
|
my $bottom_quality = $1; |
|
1829
|
|
|
|
|
|
|
# print("$bottom_quality\n"); |
|
1830
|
|
|
|
|
|
|
# print("top_quality: $top_quality bottom quality: $bottom_quality\n"); |
|
1831
|
0
|
0
|
|
|
|
|
if ($bottom_quality > $top_quality) { |
|
1832
|
|
|
|
|
|
|
# print("Chose to take the bottom quality: $bottom_quality\n"); |
|
1833
|
0
|
|
|
|
|
|
$quality = $bottom_quality; |
|
1834
|
|
|
|
|
|
|
} else { |
|
1835
|
|
|
|
|
|
|
# print("Chose to take the top quality: $top_quality\n"); |
|
1836
|
0
|
|
|
|
|
|
$quality = $top_quality; |
|
1837
|
|
|
|
|
|
|
} |
|
1838
|
|
|
|
|
|
|
} |
|
1839
|
0
|
0
|
|
|
|
|
if (!$quality) { |
|
1840
|
|
|
|
|
|
|
# print("Warning: no quality value for $current_contig, position $count!\n"); |
|
1841
|
|
|
|
|
|
|
# print("Additional data: top quality phred: $top_quality_here\n"); |
|
1842
|
|
|
|
|
|
|
# print("Additional data: bottom quality phred: $bottom_quality_here\n"); |
|
1843
|
|
|
|
|
|
|
} else { |
|
1844
|
0
|
|
|
|
|
|
$qualities[$count] = $quality; |
|
1845
|
|
|
|
|
|
|
} |
|
1846
|
|
|
|
|
|
|
} |
|
1847
|
|
|
|
|
|
|
} |
|
1848
|
|
|
|
|
|
|
|
|
1849
|
|
|
|
|
|
|
} |
|
1850
|
0
|
0
|
|
|
|
|
unless (!@qualities) { |
|
1851
|
0
|
|
|
|
|
|
$self->{'contigs'}->{$current_contig}->{'quality'} = join(" ",@qualities); |
|
1852
|
|
|
|
|
|
|
} |
|
1853
|
0
|
|
|
|
|
|
$self->{'contigs'}->{$current_contig}->{'bottom_phreds'} = undef; |
|
1854
|
0
|
|
|
|
|
|
$self->{'contigs'}->{$current_contig}->{'top_phreds'} = undef; |
|
1855
|
0
|
|
|
|
|
|
my $count = 1; |
|
1856
|
|
|
|
|
|
|
} # end foreach key |
|
1857
|
|
|
|
|
|
|
} |
|
1858
|
|
|
|
|
|
|
} |
|
1859
|
|
|
|
|
|
|
|
|
1860
|
|
|
|
|
|
|
|
|
1861
|
|
|
|
|
|
|
1; |