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# Bio::Tools::Alignment::Trim.pm |
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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::Trim - A kludge to do specialized trimming of |
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sequence based on quality. |
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=head1 SYNOPSIS |
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use Bio::Tools::Alignment::Trim; |
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$o_trim = Bio::Tools::Alignment::Trim->new(); |
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$o_trim->set_reverse_designator("R"); |
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$o_trim->set_forward_designator("F"); |
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=head1 DESCRIPTION |
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This is a specialized module designed by Chad for Chad to trim sequences |
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based on a highly specialized list of requirements. In other words, write |
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something that will trim sequences 'just like the people in the lab would |
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do manually'. |
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I settled on a sliding-window-average style of search which is ugly and |
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slow but does _exactly_ what I want it to do. |
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Mental note: rewrite this. |
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It is very important to keep in mind the context in which this module was |
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written: strictly to support the projects for which Consed.pm was |
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designed. |
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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 |
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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 bioinformatics-at-dieselwurks.com |
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=head1 APPENDIX |
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The rest of the documentation details each of the object methods. |
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Internal methods are usually preceded with a _ |
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=cut |
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package Bio::Tools::Alignment::Trim; |
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use strict; |
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use Dumpvalue; |
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use vars qw(%DEFAULTS); |
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use base qw(Bio::Root::Root); |
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BEGIN { |
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%DEFAULTS = ( 'f_designator' => 'f', |
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'r_designator' => 'r', |
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'windowsize' => '10', |
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'phreds' => '20'); |
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} |
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=head2 new() |
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Title : new() |
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Usage : $o_trim = Bio::Tools::Alignment::Trim->new(); |
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Function: Construct the Bio::Tools::Alignment::Trim object. No parameters |
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are required to create this object. It is strictly a bundle of |
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functions, as far as I am concerned. |
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Returns : A reference to a Bio::Tools::Alignment::Trim object. |
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Args : (optional) |
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-windowsize (default 10) |
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-phreds (default 20) |
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=cut |
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sub new { |
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my ($class,@args) = @_; |
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my $self = $class->SUPER::new(@args); |
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my($windowsize,$phreds) = |
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$self->_rearrange([qw( |
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WINDOWSIZE |
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PHREDS |
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)], |
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@args); |
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$self->{windowsize} = $windowsize || $DEFAULTS{'windowsize'}; |
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$self->{phreds} = $phreds || $DEFAULTS{'phreds'}; |
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# print("Constructor set phreds to ".$self->{phreds}."\n") if $self->verbose > 0; |
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$self->set_designators($DEFAULTS{'f_designator'}, |
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$DEFAULTS{'r_designator'}); |
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return $self; |
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} |
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=head2 set_designators($forward_designator,$reverse_designator) |
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Title : set_designators(,) |
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Usage : $o_trim->set_designators("F","R") |
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Function: Set the string by which the system determines whether a given |
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sequence represents a forward or a reverse read. |
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Returns : Nothing. |
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Args : two scalars: one representing the forward designator and one |
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representing the reverse designator |
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=cut |
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sub set_designators { |
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my $self = shift; |
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($self->{'f_designator'},$self->{'r_designator'}) = @_; |
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} |
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=head2 set_forward_designator($designator) |
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Title : set_forward_designator($designator) |
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Usage : $o_trim->set_forward_designator("F") |
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Function: Set the string by which the system determines if a given |
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sequence is a forward read. |
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Returns : Nothing. |
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Args : A string representing the forward designator of this project. |
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=cut |
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sub set_forward_designator { |
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my ($self,$desig) = @_; |
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$self->{'f_designator'} = $desig; |
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} |
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=head2 set_reverse_designator($reverse_designator) |
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Title : set_reverse_designator($reverse_designator) |
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Function: Set the string by which the system determines if a given |
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sequence is a reverse read. |
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Usage : $o_trim->set_reverse_designator("R") |
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Returns : Nothing. |
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Args : A string representing the forward designator of this project. |
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174
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=cut |
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sub set_reverse_designator { |
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my ($self,$desig) = @_; |
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$self->{'r_designator'} = $desig; |
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} |
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181
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=head2 get_designators() |
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183
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Title : get_designators() |
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Usage : $o_trim->get_designators() |
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Returns : A string describing the current designators. |
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Args : None |
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Notes : Really for informational purposes only. Duh. |
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189
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=cut |
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191
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sub get_designators { |
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0
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0
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1
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my $self = shift; |
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193
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0
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return("forward: ".$self->{'f_designator'}." reverse: ".$self->{'r_designator'}); |
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194
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} |
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195
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196
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=head2 trim_leading_polys() |
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197
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198
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Title : trim_leading_polys() |
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Usage : $o_trim->trim_leading_polys() |
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Function: Not implemented. Does nothing. |
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201
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Returns : Nothing. |
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202
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Args : None. |
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203
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Notes : This function is not implemented. Part of something I wanted to |
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do but never got around to doing. |
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206
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=cut |
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208
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sub trim_leading_polys { |
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209
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my ($self, $sequence) = @_; |
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} |
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211
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212
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=head2 dump_hash() |
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213
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214
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Title : dump_hash() |
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215
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Usage : $o_trim->dump_hash() |
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Function: Unimplemented. |
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217
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Returns : Nothing. |
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218
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Args : None. |
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219
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Notes : Does nothing. |
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221
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=cut |
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222
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223
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sub dump_hash { |
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224
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0
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0
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1
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my $self = shift; |
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225
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0
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my %hash = %{$self->{'qualities'}}; |
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0
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226
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} # end dump_hash |
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227
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228
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=head2 trim_singlet($sequence,$quality,$name,$class) |
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229
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230
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Title : trim_singlet($sequence,$quality,$name,$class) |
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231
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Usage : ($r_trim_points,$trimmed_sequence) = |
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232
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@{$o_trim->trim_singlet($sequence,$quality,$name,$class)}; |
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233
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Function: Trim a singlet based on its quality. |
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Returns : a reference to an array containing the forward and reverse |
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235
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trim points and the trimmed sequence. |
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236
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Args : $sequence : A sequence (SCALAR, please) |
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237
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$quality : A _scalar_ of space-delimited quality values. |
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238
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$name : the name of the sequence |
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239
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$class : The class of the sequence. One of qw(singlet |
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240
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singleton doublet pair multiplet) |
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241
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Notes : At the time this was written the bioperl objects SeqWithQuality |
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242
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and PrimaryQual did not exist. This is what is with the clumsy |
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243
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passing of references and so on. I will rewrite this next time I |
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244
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have to work with it. I also wasn't sure whether this function |
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245
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should return just the trim points or the points and the sequence. |
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246
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I decided that I always wanted both so that's how I implemented |
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247
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it. |
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248
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- Note that the size of the sliding windows is set during construction of |
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249
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the Bio::Tools::Alignment::Trim object. |
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250
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251
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=cut |
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252
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253
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sub trim_singlet { |
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254
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0
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0
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1
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my ($self,$sequence,$quality,$name,$class) = @_; |
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255
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# this split is done because I normally store quality values in a |
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256
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# space-delimited scalar rather then in an array. |
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257
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# I do this because serialization of the arrays is tough. |
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258
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0
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my @qual = split(' ',$quality); |
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259
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0
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my @points; |
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260
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0
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my $sequence_length = length($sequence); |
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261
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0
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my ($returnstring,$processed_sequence); |
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262
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# smooth out the qualities |
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263
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0
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my $r_windows = &_sliding_window(\@qual,$self->{windowsize}); |
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264
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# find out the leading and trailing trimpoints |
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265
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0
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my $start_base = $self->_get_start($r_windows,$self->{windowsize},$self->{phreds}); |
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266
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0
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my (@new_points,$trimmed_sequence); |
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267
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# do you think that any sequence shorter then 100 should be |
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268
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# discarded? I don't think that this should be the decision of this |
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269
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# module. |
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270
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# removed, 020926 |
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271
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0
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$points[0] = $start_base; |
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272
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# whew! now for the end base |
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273
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# required parameters: reference_to_windows,windowsize,$phredvalue,start_base |
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274
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my $end_base = &_get_end($r_windows,$self->{windowsize}, |
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275
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0
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|
$self->{phreds},$start_base); |
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276
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0
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|
$points[1] = $end_base; |
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277
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|
# now do the actual trimming |
|
278
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# CHAD : I don't think that it is a good idea to call chop_sequence here |
|
279
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# because chop_sequence also removes X's and N's and things |
|
280
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# and that is not always what is wanted |
|
281
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0
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|
return \@points; |
|
282
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|
} |
|
283
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|
284
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|
=head2 trim_doublet($sequence,$quality,$name,$class) |
|
285
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|
286
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|
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|
|
Title : trim_doublet($sequence,$quality,$name,$class) |
|
287
|
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|
|
Usage : ($r_trim_points,$trimmed_sequence) = |
|
288
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|
|
@{$o_trim->trim_singlet($sequence,$quality,$name,$class)}; |
|
289
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|
|
Function: Trim a singlet based on its quality. |
|
290
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|
|
Returns : a reference to an array containing the forward and reverse |
|
291
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|
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|
|
Args : $sequence : A sequence |
|
292
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|
|
$quality : A _scalar_ of space-delimited quality values. |
|
293
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|
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|
|
$name : the name of the sequence |
|
294
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|
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|
|
$class : The class of the sequence. One of qw(singlet |
|
295
|
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|
|
singleton doublet pair multiplet) |
|
296
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|
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|
|
|
Notes : At the time this was written the bioperl objects SeqWithQuality |
|
297
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|
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|
|
and PrimaryQual did not exist. This is what is with the clumsy |
|
298
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|
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|
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|
|
passing of references and so on. I will rewrite this next time I |
|
299
|
|
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|
|
|
|
have to work with it. I also wasn't sure whether this function |
|
300
|
|
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|
|
|
|
should return just the trim points or the points and the sequence. |
|
301
|
|
|
|
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|
|
I decided that I always wanted both so that's how I implemented |
|
302
|
|
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|
|
it. |
|
303
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|
304
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|
|
=cut |
|
305
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|
306
|
|
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|
#' |
|
307
|
|
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|
|
sub trim_doublet { |
|
308
|
0
|
|
|
0
|
1
|
|
my ($self,$sequence,$quality,$name,$class) = @_; |
|
309
|
0
|
|
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|
|
my @qual = split(' ',$quality); |
|
310
|
0
|
|
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|
|
my @points; |
|
311
|
0
|
|
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|
|
my $sequence_length = length($sequence); |
|
312
|
0
|
|
|
|
|
|
my ($returnstring,$processed_sequence); |
|
313
|
|
|
|
|
|
|
# smooth out the qualities |
|
314
|
0
|
|
|
|
|
|
my $r_windows = &_sliding_window(\@qual,$self->{windowsize}); |
|
315
|
|
|
|
|
|
|
# determine where the consensus sequence starts |
|
316
|
0
|
|
|
|
|
|
my $offset = 0; |
|
317
|
0
|
|
|
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|
|
for (my $current = 0; $current<$sequence_length;$current++) { |
|
318
|
0
|
0
|
|
|
|
|
if ($qual[$current] != 0) { |
|
319
|
0
|
|
|
|
|
|
$offset = $current; |
|
320
|
0
|
|
|
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|
|
last; |
|
321
|
|
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|
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|
|
} |
|
322
|
|
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|
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|
|
} |
|
323
|
|
|
|
|
|
|
# start_base required: r_quality,$windowsize,$phredvalue |
|
324
|
0
|
|
|
|
|
|
my $start_base = $self->_get_start($r_windows,$self->{windowsize},$self->{phreds},$offset); |
|
325
|
0
|
0
|
|
|
|
|
if ($start_base > ($sequence_length - 100)) { |
|
326
|
0
|
|
|
|
|
|
$points[0] = ("FAILED"); |
|
327
|
0
|
|
|
|
|
|
$points[1] = ("FAILED"); |
|
328
|
0
|
|
|
|
|
|
return @points; |
|
329
|
|
|
|
|
|
|
} |
|
330
|
0
|
|
|
|
|
|
$points[0] = $start_base; |
|
331
|
|
|
|
|
|
|
# |
|
332
|
|
|
|
|
|
|
# whew! now for the end base |
|
333
|
|
|
|
|
|
|
# |
|
334
|
|
|
|
|
|
|
# required parameters: reference_to_windows,windowsize,$phredvalue,start_base |
|
335
|
|
|
|
|
|
|
# | |
|
336
|
|
|
|
|
|
|
# 010420 NOTE: We will no longer get the end base to avoid the Q/--\___/-- syndrome |
|
337
|
0
|
|
|
|
|
|
my $end_base = $sequence_length; |
|
338
|
0
|
|
|
|
|
|
my $start_of_trailing_zeros = &count_doublet_trailing_zeros(\@qual); |
|
339
|
0
|
|
|
|
|
|
$points[1] = $end_base; |
|
340
|
|
|
|
|
|
|
# CHAD : I don't think that it is a good idea to call chop_sequence here |
|
341
|
|
|
|
|
|
|
# because chop_sequence also removes X's and N's and things |
|
342
|
|
|
|
|
|
|
# and that is not always what is wanted |
|
343
|
0
|
|
|
|
|
|
return @points; |
|
344
|
|
|
|
|
|
|
} # end trim_doublet |
|
345
|
|
|
|
|
|
|
|
|
346
|
|
|
|
|
|
|
=head2 chop_sequence($name,$class,$sequence,@points) |
|
347
|
|
|
|
|
|
|
|
|
348
|
|
|
|
|
|
|
Title : chop_sequence($name,$class,$sequence,@points) |
|
349
|
|
|
|
|
|
|
Usage : ($start_point,$end_point,$chopped_sequence) = |
|
350
|
|
|
|
|
|
|
$o_trim->chop_sequence($name,$class,$sequence,@points); |
|
351
|
|
|
|
|
|
|
Function: Chop a sequence based on its name, class, and sequence. |
|
352
|
|
|
|
|
|
|
Returns : an array containing three scalars: |
|
353
|
|
|
|
|
|
|
1- the start trim point |
|
354
|
|
|
|
|
|
|
2- the end trim point |
|
355
|
|
|
|
|
|
|
3- the chopped sequence |
|
356
|
|
|
|
|
|
|
Args : |
|
357
|
|
|
|
|
|
|
$name : the name of the sequence |
|
358
|
|
|
|
|
|
|
$class : The class of the sequence. One of qw(singlet |
|
359
|
|
|
|
|
|
|
singleton doublet pair multiplet) |
|
360
|
|
|
|
|
|
|
$sequence : A sequence |
|
361
|
|
|
|
|
|
|
@points : An array containing two elements- the first contains |
|
362
|
|
|
|
|
|
|
the start trim point and the second conatines the end trim |
|
363
|
|
|
|
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|
|
point. |
|
364
|
|
|
|
|
|
|
|
|
365
|
|
|
|
|
|
|
=cut |
|
366
|
|
|
|
|
|
|
|
|
367
|
|
|
|
|
|
|
sub chop_sequence { |
|
368
|
0
|
|
|
0
|
1
|
|
my ($self,$name,$class,$sequence,@points) = @_; |
|
369
|
0
|
|
|
|
|
|
print("Coming into chop_sequence, \@points are @points\n"); |
|
370
|
0
|
|
|
|
|
|
my $fdesig = $self->{'f_designator'}; |
|
371
|
0
|
|
|
|
|
|
my $rdesig = $self->{'r_designator'}; |
|
372
|
0
|
0
|
0
|
|
|
|
if (!$points[0] && !$points[1]) { |
|
373
|
0
|
|
|
|
|
|
$sequence = "junk"; |
|
374
|
0
|
|
|
|
|
|
return $sequence; |
|
375
|
|
|
|
|
|
|
} |
|
376
|
0
|
0
|
0
|
|
|
|
if ($class eq "singlet" && $name =~ /$fdesig$/) { |
|
|
|
0
|
0
|
|
|
|
|
|
|
|
0
|
0
|
|
|
|
|
|
|
|
0
|
0
|
|
|
|
|
|
|
|
0
|
|
|
|
|
|
|
377
|
0
|
|
|
|
|
|
$sequence = substr($sequence,$points[0],$points[1]-$points[0]); |
|
378
|
|
|
|
|
|
|
} |
|
379
|
|
|
|
|
|
|
elsif ($class eq "singlet" && $name =~ /$rdesig$/) { |
|
380
|
0
|
|
|
|
|
|
$sequence = substr($sequence,$points[0],$points[1]-$points[0]); |
|
381
|
|
|
|
|
|
|
} |
|
382
|
|
|
|
|
|
|
elsif ($class eq "singleton" && $name =~ /$fdesig$/) { |
|
383
|
0
|
|
|
|
|
|
$sequence = substr($sequence,$points[0],$points[1]-$points[0]); |
|
384
|
|
|
|
|
|
|
} |
|
385
|
|
|
|
|
|
|
elsif ($class eq "singleton" && $name =~ /$rdesig$/) { |
|
386
|
0
|
|
|
|
|
|
$sequence = substr($sequence,$points[0],$points[1]-$points[0]); |
|
387
|
|
|
|
|
|
|
} |
|
388
|
|
|
|
|
|
|
elsif ($class eq "doublet") { |
|
389
|
0
|
|
|
|
|
|
$sequence = substr($sequence,$points[0],$points[1]-$points[0]); |
|
390
|
|
|
|
|
|
|
} |
|
391
|
|
|
|
|
|
|
# this is a _terrible_ to do this! i couldn't seem to find a better way |
|
392
|
|
|
|
|
|
|
# i thought something like s/(^.*[Xx]{5,})//g; might work, but no go |
|
393
|
|
|
|
|
|
|
# no time to find a fix! |
|
394
|
0
|
|
|
|
|
|
my $length_before_trimming = length($sequence); |
|
395
|
0
|
|
|
|
|
|
my $subs_Xs = $sequence =~ s/^.*[Xx]{5,}//g; |
|
396
|
0
|
0
|
|
|
|
|
if ($subs_Xs) { |
|
397
|
0
|
|
|
|
|
|
my $length_after_trimming = length($sequence); |
|
398
|
0
|
|
|
|
|
|
my $number_Xs_trimmed = $length_before_trimming - $length_after_trimming; |
|
399
|
0
|
|
|
|
|
|
$points[0] += $number_Xs_trimmed; |
|
400
|
|
|
|
|
|
|
} |
|
401
|
0
|
|
|
|
|
|
$length_before_trimming = length($sequence); |
|
402
|
0
|
|
|
|
|
|
my $subs_Ns = $sequence =~ s/[Nn]{1,}$//g; |
|
403
|
0
|
0
|
|
|
|
|
if ($subs_Ns) { |
|
404
|
0
|
|
|
|
|
|
my $length_after_trimming = length($sequence); |
|
405
|
0
|
|
|
|
|
|
my $number_Ns_trimmed = $length_before_trimming - $length_after_trimming; |
|
406
|
0
|
|
|
|
|
|
$points[1] -= $number_Ns_trimmed; |
|
407
|
0
|
|
|
|
|
|
$points[1] -= 1; |
|
408
|
|
|
|
|
|
|
} |
|
409
|
0
|
|
|
|
|
|
push @points,$sequence; |
|
410
|
0
|
|
|
|
|
|
print("chop_sequence \@points are @points\n"); |
|
411
|
0
|
|
|
|
|
|
return @points; |
|
412
|
|
|
|
|
|
|
} |
|
413
|
|
|
|
|
|
|
|
|
414
|
|
|
|
|
|
|
=head2 _get_start($r_quals,$windowsize,$phreds,$offset) |
|
415
|
|
|
|
|
|
|
|
|
416
|
|
|
|
|
|
|
Title : _get_start($r_quals,$windowsize,$phreds,$offset) |
|
417
|
|
|
|
|
|
|
Usage : $start_base = $self->_get_start($r_windows,5,20); |
|
418
|
|
|
|
|
|
|
Function: Provide the start trim point for this sequence. |
|
419
|
|
|
|
|
|
|
Returns : a scalar representing the start of the sequence |
|
420
|
|
|
|
|
|
|
Args : |
|
421
|
|
|
|
|
|
|
$r_quals : A reference to an array containing quality values. In |
|
422
|
|
|
|
|
|
|
context, this array of values has been smoothed by then |
|
423
|
|
|
|
|
|
|
sliding window-look ahead algorithm. |
|
424
|
|
|
|
|
|
|
$windowsize : The size of the window used when the sliding window |
|
425
|
|
|
|
|
|
|
look-ahead average was calculated. |
|
426
|
|
|
|
|
|
|
$phreds : |
|
427
|
|
|
|
|
|
|
$offset : |
|
428
|
|
|
|
|
|
|
|
|
429
|
|
|
|
|
|
|
=cut |
|
430
|
|
|
|
|
|
|
|
|
431
|
|
|
|
|
|
|
sub _get_start { |
|
432
|
0
|
|
|
0
|
|
|
my ($self,$r_quals,$windowsize,$phreds,$offset) = @_; |
|
433
|
0
|
0
|
|
|
|
|
print("Using $phreds phreds\n") if $self->verbose > 0; |
|
434
|
|
|
|
|
|
|
# this is to help determine whether the sequence is good at all |
|
435
|
0
|
|
|
|
|
|
my @quals = @$r_quals; |
|
436
|
0
|
|
|
|
|
|
my ($count,$count2,$qualsum); |
|
437
|
0
|
0
|
|
|
|
|
if ($offset) { $count = $offset; } else { $count = 0; } |
|
|
0
|
|
|
|
|
|
|
|
|
0
|
|
|
|
|
|
|
|
438
|
|
|
|
|
|
|
# search along the length of the sequence |
|
439
|
0
|
|
|
|
|
|
for (; ($count+$windowsize) <= scalar(@quals); $count++) { |
|
440
|
|
|
|
|
|
|
# sum all of the quality values in this window. |
|
441
|
0
|
|
|
|
|
|
my $cumulative=0; |
|
442
|
0
|
|
|
|
|
|
for($count2 = $count; $count2 < $count+$windowsize; $count2++) { |
|
443
|
0
|
0
|
|
|
|
|
if (!$quals[$count2]) { |
|
444
|
|
|
|
|
|
|
# print("Quals don't exist here!\n"); |
|
445
|
|
|
|
|
|
|
} |
|
446
|
|
|
|
|
|
|
else { |
|
447
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
448
|
|
|
|
|
|
|
# print("Incremented qualsum to ($qualsum)\n"); |
|
449
|
|
|
|
|
|
|
} |
|
450
|
0
|
|
|
|
|
|
$cumulative++; |
|
451
|
|
|
|
|
|
|
} |
|
452
|
|
|
|
|
|
|
# print("The sum of this window (starting at $count) is $qualsum. I counted $cumulative bases.\n"); |
|
453
|
|
|
|
|
|
|
# if the total of windowsize * phreds is |
|
454
|
0
|
0
|
0
|
|
|
|
if ($qualsum && $qualsum >= $windowsize*$phreds) { return $count; } |
|
|
0
|
|
|
|
|
|
|
|
455
|
0
|
|
|
|
|
|
$qualsum = 0; |
|
456
|
|
|
|
|
|
|
} |
|
457
|
|
|
|
|
|
|
# if ($count > scalar(@quals)-$windowsize) { return; } |
|
458
|
0
|
|
|
|
|
|
return $count; |
|
459
|
|
|
|
|
|
|
} |
|
460
|
|
|
|
|
|
|
|
|
461
|
|
|
|
|
|
|
=head2 _get_end($r_qual,$windowsize,$phreds,$count) |
|
462
|
|
|
|
|
|
|
|
|
463
|
|
|
|
|
|
|
Title : _get_end($r_qual,$windowsize,$phreds,$count) |
|
464
|
|
|
|
|
|
|
Usage : my $end_base = &_get_end($r_windows,20,20,$start_base); |
|
465
|
|
|
|
|
|
|
Function: Get the end trim point for this sequence. |
|
466
|
|
|
|
|
|
|
Returns : A scalar representing the end trim point for this sequence. |
|
467
|
|
|
|
|
|
|
Args : |
|
468
|
|
|
|
|
|
|
$r_qual : A reference to an array containing quality values. In |
|
469
|
|
|
|
|
|
|
context, this array of values has been smoothed by then |
|
470
|
|
|
|
|
|
|
sliding window-look ahead algorithm. |
|
471
|
|
|
|
|
|
|
$windowsize : The size of the window used when the sliding window |
|
472
|
|
|
|
|
|
|
look-ahead average was calculated. |
|
473
|
|
|
|
|
|
|
$phreds : |
|
474
|
|
|
|
|
|
|
$count : Start looking for the end of the sequence here. |
|
475
|
|
|
|
|
|
|
|
|
476
|
|
|
|
|
|
|
=cut |
|
477
|
|
|
|
|
|
|
|
|
478
|
|
|
|
|
|
|
sub _get_end { |
|
479
|
0
|
|
|
0
|
|
|
my ($r_qual,$windowsize,$phreds,$count) = @_; |
|
480
|
0
|
|
|
|
|
|
my @quals = @$r_qual; |
|
481
|
0
|
|
|
|
|
|
my $total_bases = scalar(@quals); |
|
482
|
0
|
|
|
|
|
|
my ($count2,$qualsum,$end_of_quals,$bases_counted); |
|
483
|
0
|
0
|
|
|
|
|
if (!$count) { $count=0; } |
|
|
0
|
|
|
|
|
|
|
|
484
|
0
|
|
|
|
|
|
BASE: for (; $count < $total_bases; $count++) { |
|
485
|
0
|
|
|
|
|
|
$bases_counted = 0; |
|
486
|
0
|
|
|
|
|
|
$qualsum = 0; |
|
487
|
0
|
|
|
|
|
|
POSITION: for($count2 = $count; $count2 < $total_bases; $count2++) { |
|
488
|
0
|
|
|
|
|
|
$bases_counted++; |
|
489
|
|
|
|
|
|
|
|
|
490
|
0
|
0
|
|
|
|
|
if ($count2 == $total_bases-1) { |
|
|
|
0
|
|
|
|
|
|
|
491
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
492
|
0
|
|
|
|
|
|
$bases_counted++; |
|
493
|
0
|
|
|
|
|
|
last BASE; |
|
494
|
|
|
|
|
|
|
} |
|
495
|
|
|
|
|
|
|
elsif ($bases_counted == $windowsize) { |
|
496
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
497
|
0
|
0
|
|
|
|
|
if ($qualsum < $bases_counted*$phreds) { |
|
498
|
0
|
|
|
|
|
|
return $count+$bases_counted+$windowsize; |
|
499
|
|
|
|
|
|
|
} |
|
500
|
0
|
|
|
|
|
|
next BASE; |
|
501
|
|
|
|
|
|
|
} |
|
502
|
|
|
|
|
|
|
else { |
|
503
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
504
|
|
|
|
|
|
|
} |
|
505
|
|
|
|
|
|
|
} |
|
506
|
0
|
0
|
|
|
|
|
if ($qualsum < $bases_counted*$phreds) { |
|
507
|
0
|
|
|
|
|
|
return $count+$bases_counted+$windowsize; |
|
508
|
|
|
|
|
|
|
} |
|
509
|
|
|
|
|
|
|
else { } |
|
510
|
0
|
|
|
|
|
|
$qualsum = 0; |
|
511
|
|
|
|
|
|
|
} # end for |
|
512
|
0
|
0
|
|
|
|
|
if ($end_of_quals) { |
|
513
|
0
|
|
|
|
|
|
my $bases_for_average = $total_bases-$count2; |
|
514
|
0
|
|
|
|
|
|
return $count2; |
|
515
|
|
|
|
|
|
|
} |
|
516
|
|
|
|
|
|
|
else { } |
|
517
|
0
|
0
|
|
|
|
|
if ($qualsum) { } # print ("$qualsum\n"); |
|
518
|
0
|
|
|
|
|
|
return $total_bases; |
|
519
|
|
|
|
|
|
|
} # end get_end |
|
520
|
|
|
|
|
|
|
|
|
521
|
|
|
|
|
|
|
=head2 count_doublet_trailing_zeros($r_qual) |
|
522
|
|
|
|
|
|
|
|
|
523
|
|
|
|
|
|
|
Title : count_doublet_trailing_zeros($r_qual) |
|
524
|
|
|
|
|
|
|
Usage : my $start_of_trailing_zeros = &count_doublet_trailing_zeros(\@qual); |
|
525
|
|
|
|
|
|
|
Function: Find out when the trailing zero qualities start. |
|
526
|
|
|
|
|
|
|
Returns : A scalar representing where the zeros start. |
|
527
|
|
|
|
|
|
|
Args : A reference to an array of quality values. |
|
528
|
|
|
|
|
|
|
Notes : Again, this should be rewritten to use PrimaryQual objects. |
|
529
|
|
|
|
|
|
|
A more detailed explanation of why phrap puts these zeros here should |
|
530
|
|
|
|
|
|
|
be written and placed here. Please email and hassle the author. |
|
531
|
|
|
|
|
|
|
|
|
532
|
|
|
|
|
|
|
|
|
533
|
|
|
|
|
|
|
=cut |
|
534
|
|
|
|
|
|
|
|
|
535
|
|
|
|
|
|
|
sub count_doublet_trailing_zeros { |
|
536
|
0
|
|
|
0
|
1
|
|
my ($r_qual) = shift; |
|
537
|
0
|
|
|
|
|
|
my $number_of_trailing_zeros = 0; |
|
538
|
0
|
|
|
|
|
|
my @qualities = @$r_qual; |
|
539
|
0
|
|
|
|
|
|
for (my $current=scalar(@qualities);$current>0;$current--) { |
|
540
|
0
|
0
|
0
|
|
|
|
if ($qualities[$current] && $qualities[$current] != 0) { |
|
541
|
0
|
|
|
|
|
|
$number_of_trailing_zeros = scalar(@qualities)-$current; |
|
542
|
0
|
|
|
|
|
|
return $current+1; |
|
543
|
|
|
|
|
|
|
} |
|
544
|
|
|
|
|
|
|
} |
|
545
|
0
|
|
|
|
|
|
return scalar(@qualities); |
|
546
|
|
|
|
|
|
|
} # end count_doublet_trailing_zeros |
|
547
|
|
|
|
|
|
|
|
|
548
|
|
|
|
|
|
|
=head2 _sliding_window($r_quals,$windowsize) |
|
549
|
|
|
|
|
|
|
|
|
550
|
|
|
|
|
|
|
Title : _sliding_window($r_quals,$windowsize) |
|
551
|
|
|
|
|
|
|
Usage : my $r_windows = &_sliding_window(\@qual,$windowsize); |
|
552
|
|
|
|
|
|
|
Function: Create a sliding window, look-forward-average on an array |
|
553
|
|
|
|
|
|
|
of quality values. Used to smooth out differences in qualities. |
|
554
|
|
|
|
|
|
|
Returns : A reference to an array containing the smoothed values. |
|
555
|
|
|
|
|
|
|
Args : $r_quals: A reference to an array containing quality values. |
|
556
|
|
|
|
|
|
|
$windowsize : The size of the sliding window. |
|
557
|
|
|
|
|
|
|
Notes : This was written before PrimaryQual objects existed. They |
|
558
|
|
|
|
|
|
|
should use that object but I haven't rewritten this yet. |
|
559
|
|
|
|
|
|
|
|
|
560
|
|
|
|
|
|
|
=cut |
|
561
|
|
|
|
|
|
|
|
|
562
|
|
|
|
|
|
|
#' |
|
563
|
|
|
|
|
|
|
sub _sliding_window { |
|
564
|
0
|
|
|
0
|
|
|
my ($r_quals,$windowsize) = @_; |
|
565
|
0
|
|
|
|
|
|
my (@window,@quals,$qualsum,$count,$count2,$average,@averages,$bases_counted); |
|
566
|
0
|
|
|
|
|
|
@quals = @$r_quals; |
|
567
|
0
|
|
|
|
|
|
my $size_of_quality = scalar(@quals); |
|
568
|
|
|
|
|
|
|
# do this loop for all of the qualities |
|
569
|
0
|
|
|
|
|
|
for ($count=0; $count <= $size_of_quality; $count++) { |
|
570
|
0
|
|
|
|
|
|
$bases_counted = 0; |
|
571
|
0
|
|
|
|
|
|
BASE: for($count2 = $count; $count2 < $size_of_quality; $count2++) { |
|
572
|
0
|
|
|
|
|
|
$bases_counted++; |
|
573
|
|
|
|
|
|
|
# if the search hits the end of the averages, stop |
|
574
|
|
|
|
|
|
|
# this is for the case near the end where bases remaining < windowsize |
|
575
|
0
|
0
|
|
|
|
|
if ($count2 == $size_of_quality) { |
|
|
|
0
|
|
|
|
|
|
|
576
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
577
|
0
|
|
|
|
|
|
last BASE; |
|
578
|
|
|
|
|
|
|
} |
|
579
|
|
|
|
|
|
|
# if the search hits the size of the window |
|
580
|
|
|
|
|
|
|
elsif ($bases_counted == $windowsize) { |
|
581
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
582
|
0
|
|
|
|
|
|
last BASE; |
|
583
|
|
|
|
|
|
|
} |
|
584
|
|
|
|
|
|
|
# otherwise add the quality value |
|
585
|
0
|
0
|
|
|
|
|
unless (!$quals[$count2]) { |
|
586
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
587
|
|
|
|
|
|
|
} |
|
588
|
|
|
|
|
|
|
} |
|
589
|
0
|
0
|
0
|
|
|
|
unless (!$qualsum || !$windowsize) { |
|
590
|
0
|
|
|
|
|
|
$average = $qualsum / $bases_counted; |
|
591
|
0
|
0
|
|
|
|
|
if (!$average) { $average = "0"; } |
|
|
0
|
|
|
|
|
|
|
|
592
|
0
|
|
|
|
|
|
push @averages,$average; |
|
593
|
|
|
|
|
|
|
} |
|
594
|
0
|
|
|
|
|
|
$qualsum = 0; |
|
595
|
|
|
|
|
|
|
} |
|
596
|
|
|
|
|
|
|
# 02101 Yes, I repaired the mismatching numbers between averages and windows. |
|
597
|
|
|
|
|
|
|
# print("There are ".scalar(@$r_quals)." quality values. They are @$r_quals\n"); |
|
598
|
|
|
|
|
|
|
# print("There are ".scalar(@averages)." average values. They are @averages\n"); |
|
599
|
0
|
|
|
|
|
|
return \@averages; |
|
600
|
|
|
|
|
|
|
|
|
601
|
|
|
|
|
|
|
} |
|
602
|
|
|
|
|
|
|
|
|
603
|
|
|
|
|
|
|
=head2 _print_formatted_qualities |
|
604
|
|
|
|
|
|
|
|
|
605
|
|
|
|
|
|
|
Title : _print_formatted_qualities(\@quals) |
|
606
|
|
|
|
|
|
|
Usage : &_print_formatted_qualities(\@quals); |
|
607
|
|
|
|
|
|
|
Returns : Nothing. Prints. |
|
608
|
|
|
|
|
|
|
Args : A reference to an array containing quality values. |
|
609
|
|
|
|
|
|
|
Notes : An internal procedure used in debugging. Prints out an array nicely. |
|
610
|
|
|
|
|
|
|
|
|
611
|
|
|
|
|
|
|
=cut |
|
612
|
|
|
|
|
|
|
|
|
613
|
|
|
|
|
|
|
sub _print_formatted_qualities { |
|
614
|
0
|
|
|
0
|
|
|
my $rquals = shift; |
|
615
|
0
|
|
|
|
|
|
my @qual = @$rquals; |
|
616
|
0
|
|
|
|
|
|
for (my $count=0; $count
|
|
617
|
0
|
0
|
|
|
|
|
if (($count%10)==0) { print("\n$count\t"); } |
|
|
0
|
|
|
|
|
|
|
|
618
|
0
|
0
|
|
|
|
|
if ($qual[$count]) { print ("$qual[$count]\t");} |
|
|
0
|
|
|
|
|
|
|
|
619
|
0
|
|
|
|
|
|
else { print("0\t"); } |
|
620
|
|
|
|
|
|
|
} |
|
621
|
0
|
|
|
|
|
|
print("\n"); |
|
622
|
|
|
|
|
|
|
} |
|
623
|
|
|
|
|
|
|
|
|
624
|
|
|
|
|
|
|
=head2 _get_end_old($r_qual,$windowsize,$phreds,$count) |
|
625
|
|
|
|
|
|
|
|
|
626
|
|
|
|
|
|
|
Title : _get_end_old($r_qual,$windowsize,$phreds,$count) |
|
627
|
|
|
|
|
|
|
Usage : Deprecated. Don't use this! |
|
628
|
|
|
|
|
|
|
Returns : Deprecated. Don't use this! |
|
629
|
|
|
|
|
|
|
Args : Deprecated. Don't use this! |
|
630
|
|
|
|
|
|
|
|
|
631
|
|
|
|
|
|
|
=cut |
|
632
|
|
|
|
|
|
|
|
|
633
|
|
|
|
|
|
|
#' |
|
634
|
|
|
|
|
|
|
sub _get_end_old { |
|
635
|
0
|
|
|
0
|
|
|
my ($r_qual,$windowsize,$phreds,$count) = @_; |
|
636
|
0
|
|
|
|
|
|
warn("Do Not Use this function (_get_end_old)"); |
|
637
|
0
|
|
|
|
|
|
my $target = $windowsize*$phreds; |
|
638
|
0
|
|
|
|
|
|
my @quals = @$r_qual; |
|
639
|
0
|
|
|
|
|
|
my $total_bases = scalar(@quals); |
|
640
|
0
|
|
|
|
|
|
my ($count2,$qualsum,$end_of_quals); |
|
641
|
0
|
0
|
|
|
|
|
if (!$count) { $count=0; } |
|
|
0
|
|
|
|
|
|
|
|
642
|
0
|
|
|
|
|
|
BASE: for (; $count < $total_bases; $count++) { |
|
643
|
0
|
|
|
|
|
|
for($count2 = $count; $count2 < $count+$windowsize; $count2++) { |
|
644
|
0
|
0
|
|
|
|
|
if ($count2 == scalar(@quals)-1) { |
|
645
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
646
|
0
|
|
|
|
|
|
$end_of_quals = 1; |
|
647
|
0
|
|
|
|
|
|
last BASE; |
|
648
|
|
|
|
|
|
|
|
|
649
|
|
|
|
|
|
|
} |
|
650
|
0
|
|
|
|
|
|
$qualsum += $quals[$count2]; |
|
651
|
|
|
|
|
|
|
} |
|
652
|
0
|
0
|
|
|
|
|
if ($qualsum < $windowsize*$phreds) { |
|
653
|
0
|
|
|
|
|
|
return $count+$windowsize; |
|
654
|
|
|
|
|
|
|
} |
|
655
|
0
|
|
|
|
|
|
$qualsum = 0; |
|
656
|
|
|
|
|
|
|
} # end for |
|
657
|
|
|
|
|
|
|
} # end get_end_old |
|
658
|
|
|
|
|
|
|
|
|
659
|
|
|
|
|
|
|
|
|
660
|
|
|
|
|
|
|
# Autoload methods go after =cut, and are processed by the autosplit program. |
|
661
|
|
|
|
|
|
|
|
|
662
|
|
|
|
|
|
|
1; |
|
663
|
|
|
|
|
|
|
__END__ |