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# BioPerl module for Bio::Tools::Run::Newbler |
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# |
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# Copyright Florent E Angly |
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# |
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# You may distribute this module under the same terms as perl itself |
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# |
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# POD documentation - main docs before the code |
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=head1 NAME |
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Bio::Tools::Run::Newbler - Wrapper for local execution of Newbler |
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=head1 SYNOPSIS |
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use Bio::Tools::Run::Newbler; |
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# Run Minmo using an input FASTA file |
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my $factory = Bio::Tools::Run::Newbler->new( -minimum_overlap_length => 35 ); |
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my $asm_obj = $factory->run($fasta_file, $qual_file); |
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# An assembly object is returned by default |
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for my $contig ($assembly->all_contigs) { |
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... do something ... |
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} |
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# Read some sequences |
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use Bio::SeqIO; |
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my $sio = Bio::SeqIO->new(-file => $fasta_file, -format => 'fasta'); |
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my @seqs; |
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while (my $seq = $sio->next_seq()) { |
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push @seqs,$seq; |
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} |
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# Run Newbler using input sequence objects and returning an assembly file |
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my $asm_file = 'results.ace'; |
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$factory->out_type($asm_file); |
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$factory->run(\@seqs); |
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=head1 DESCRIPTION |
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Wrapper module for the local execution of the proprietary DNA assembly |
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program GS De Novo Assembler (Newbler) from Roche/454 v2.0.00.20: |
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http://www.454.com/products-solutions/analysis-tools/gs-de-novo-assembler.asp |
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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 Bioperl |
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modules. Send your comments and suggestions preferably to one of the Bioperl |
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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 the bugs |
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and their resolution. Bug reports can be submitted via the web: |
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http://redmine.open-bio.org/projects/bioperl/ |
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=head1 AUTHOR - Florent E Angly |
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Email: florent-dot-angly-at-gmail-dot-com |
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=head1 APPENDIX |
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The rest of the documentation details each of the object methods. Internal |
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methods are usually preceded with a _ |
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=cut |
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package Bio::Tools::Run::Newbler; |
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use strict; |
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use IPC::Run; |
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use File::Copy; |
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use File::Path; |
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use File::Spec; |
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use File::Basename; |
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use base qw( Bio::Root::Root Bio::Tools::Run::AssemblerBase ); |
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our $program_name = 'runAssembly'; # name of the executable |
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our @program_params = (qw( expected_depth mid_conf_file vector_trim vector_screen |
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aln_identity_score aln_difference_score min_ovl_identity min_ovl_length |
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seed_count seed_length seed_step out_dir )); |
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our @program_switches = (qw( large ace ace_raw ace_trimmed no_trim in_memory |
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no_auto_rescore no_duplicates )); |
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our %param_translation = ( |
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'large' => 'large', |
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'ace' => 'ace', |
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'ace_raw' => 'ar', |
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'ace_trimmed' => 'at', |
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'expected_depth' => 'e', |
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'mid_conf_file' => 'mcf', |
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'no_trim' => 'notrim', |
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'vector_trim' => 'vt', |
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'vector_screen' => 'vs', |
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'aln_identity_score' => 'ais', |
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'aln_difference_score'=> 'ads', |
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'in_memory' => 'm', |
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'min_ovl_identity' => 'mi', |
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'min_ovl_length' => 'ml', |
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'no_auto_rescore' => 'nor', |
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'seed_count' => 'sc', |
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'seed_length' => 'sl', |
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'seed_step' => 'ss', |
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'no_duplicates' => 'ud', |
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'out_dir' => 'o' |
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); |
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our $qual_param; |
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our $use_dash = 1; |
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our $join = ' '; |
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our $asm_format = 'ace'; |
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our $asm_variant = '454'; |
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=head2 new |
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Title : new |
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Usage : $assembler->new( -min_len => 50, |
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-min_ident => 95 ); |
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Function: Creates a Newbler factory |
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Returns : A Bio::Tools::Run::Newbler object |
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Args : Newbler options available in this module (from the Newbler manual): |
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large Shortcut some of the computationally expensive algorithms |
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to save some time. Useful for large or complex datasets |
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(default: off). |
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ace_raw Output the full "raw" read sequence (default: off). |
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ace_trimmed Output only the "trimmed" sequences (after low quality, |
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vector and key trimming) (default: on). |
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expected_depth Expected depth of the assembly. Filters out random-chance |
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level events at bigger depths. 0 means to not use the |
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expected depth information (default: 0). |
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mid_conf_file MID configuration file for decoding the multiplex data. |
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no_trim Disable the quality and primer trimming of the input |
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sequences (default: off). |
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vector_trim Specify a vector trimming database (in FASTA format) to |
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trim the ends of input sequences. |
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vector_screen Specify a vector screening database (in FASTA format) to |
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remove contaminants, i.e. input reads that align |
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against a sequence in the database. |
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aln_identity_score Set the alignment identity score. When multiple alignments |
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are found, it is the per-overlap column identity score |
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used to sort the overlaps for use in the progressive |
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alignment (default: 2). |
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aln_difference_score Set the alignment difference score. For multiple alignments |
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this is the per-overlap difference score used to sort the |
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overlaps for use in the progressive multi-alignment |
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(default: -3). |
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in_memory Keep all sequence data in memory throughout the computation. |
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Can speed up the computation but requires more computer |
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memory (default: off). |
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min_ovl_identity / minimum_overlap_similarity |
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Minimum overlap identity, i.e. the minimum percent identity |
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of overlaps used by the assembler (default: 40). |
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min_ovl_length / minimum_overlap_length |
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Minimum overlap length, i.e. the minimum length of overlaps |
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considered by the assembler (default: 90). Warning: It |
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seems like this parameter is not respected by the program |
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in the current version |
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no_auto_rescore Do not use the quality score re-scoring algorithm (default: |
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off). |
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seed_count Set the seed count parameter, the number of seeds required |
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in a window before an extension is made (default: 1). |
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seed_length Set the seed length parameter, i.e. the number of bases |
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between seed generation locations used in the exact |
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k-mer matching part of the overlap detection (between 6 |
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16) (default: 16). |
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seed_step Set the seed step parameter, i.e. the number of bases used |
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for each seed in the exact k-mer matching part of the |
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overlap detection (i.e. the "k" value) (default: 12). |
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no_duplicates Treat each read as a separate read and do not group them |
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into duplicates for assembly or consensus calling |
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(default: off). |
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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->_set_program_options(\@args, \@program_params, \@program_switches, |
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\%param_translation, $qual_param, $use_dash, $join); |
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*minimum_overlap_length = \&min_ovl_length; |
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*minimum_overlap_similarity = \&min_ovl_identity; |
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$self->program_name($program_name) if not defined $self->program_name(); |
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$self->_assembly_format($asm_format); |
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$self->_assembly_variant($asm_variant); |
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return $self; |
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} |
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=head2 _check_sequence_input |
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Title : _check_sequence_input |
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Usage : $assembler->_check_sequence_input($seqs) |
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Function: Check that the sequence input is arrayref of sequence objects or |
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a FASTA file, or a MIDinfo + dir, or a MIDinfo + file. If not, an |
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error is thrown. |
213
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Returns : 1 if the check passed |
214
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Args : sequence input |
215
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216
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=cut |
217
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218
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sub _check_sequence_input { |
219
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4
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4
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463
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my ($self, $seqs) = @_; |
220
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4
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50
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9
|
if (not $seqs) { |
221
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0
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0
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$self->throw("Must supply sequences as a FASTA filename or a sequence object". |
222
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" (Bio::PrimarySeqI or Bio::SeqI) array reference"); |
223
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} else { |
224
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4
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50
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6
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if (ref($seqs) =~ m/ARRAY/i ) { |
225
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0
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0
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for my $seq (@$seqs) { |
226
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0
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0
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0
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0
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unless ($seq->isa('Bio::PrimarySeqI') || $seq->isa('Bio::SeqI')) { |
227
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0
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0
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$self->throw("Not a valid Bio::PrimarySeqI or Bio::SeqI object"); |
228
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} |
229
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} |
230
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} else { |
231
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# [midinfo@]sffile|[midinfo@]projectdir|fastafile |
232
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4
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19
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my ($mid, $file_or_dir) = ($seqs =~ m/^(.+@)?(.+)$/); |
233
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4
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50
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|
5
|
if (not defined $file_or_dir) { |
234
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0
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|
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|
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0
|
$self->throw("Input string $seqs does not seem valid."); |
235
|
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} else { |
236
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4
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50
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57
|
if (not -e $file_or_dir) { |
237
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0
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|
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0
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$self->throw("Input file or directory '$file_or_dir' does not seem to exist."); |
238
|
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} |
239
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} |
240
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} |
241
|
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} |
242
|
4
|
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13
|
return 1; |
243
|
|
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|
} |
244
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245
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|
=head2 out_type |
246
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247
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Title : out_type |
248
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|
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|
|
Usage : $factory->out_type('Bio::Assembly::ScaffoldI') |
249
|
|
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|
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|
Function: Get/set the desired type of output |
250
|
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|
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|
Returns : The type of results to return |
251
|
|
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|
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|
Args : Desired type of results to return (optional): |
252
|
|
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|
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|
|
'Bio::Assembly::IO' object |
253
|
|
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|
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|
|
'Bio::Assembly::ScaffoldI' object (default) |
254
|
|
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|
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|
|
The name of a file to save the results in |
255
|
|
|
|
|
|
|
|
256
|
|
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|
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|
|
=cut |
257
|
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|
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258
|
|
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|
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259
|
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|
|
=head2 run |
260
|
|
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|
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261
|
|
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|
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|
|
Title : run |
262
|
|
|
|
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|
|
Usage : $factory->run($fasta_file); |
263
|
|
|
|
|
|
|
Function: Run TIGR Assembler |
264
|
|
|
|
|
|
|
Returns : - a Bio::Assembly::ScaffoldI object, a Bio::Assembly::IO |
265
|
|
|
|
|
|
|
object, a filename, or undef if all sequences were too small to |
266
|
|
|
|
|
|
|
be usable |
267
|
|
|
|
|
|
|
Returns : Assembly results (file, IO object or assembly object) |
268
|
|
|
|
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|
|
Args : Sequence input can be: |
269
|
|
|
|
|
|
|
* a sequence object arrayref |
270
|
|
|
|
|
|
|
* a FASTA file |
271
|
|
|
|
|
|
|
* a SFF file and optional MID information. Example: |
272
|
|
|
|
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|
|
mid2@/home/xxx/myreads.sff |
273
|
|
|
|
|
|
|
* the path to an run analysis directory and MID information |
274
|
|
|
|
|
|
|
The reads must be between 50 and 2000 bp. Newbler does not support |
275
|
|
|
|
|
|
|
for input quality files. See the Newbler manual for details. |
276
|
|
|
|
|
|
|
|
277
|
|
|
|
|
|
|
=cut |
278
|
|
|
|
|
|
|
|
279
|
|
|
|
|
|
|
|
280
|
|
|
|
|
|
|
=head2 _run |
281
|
|
|
|
|
|
|
|
282
|
|
|
|
|
|
|
Title : _run |
283
|
|
|
|
|
|
|
Usage : $factory->_run() |
284
|
|
|
|
|
|
|
Function: Make a system call and run TIGR Assembler |
285
|
|
|
|
|
|
|
Returns : An assembly file |
286
|
|
|
|
|
|
|
Args : - FASTA file, SFF file and MID, or analysis dir and MID |
287
|
|
|
|
|
|
|
- optional QUAL file |
288
|
|
|
|
|
|
|
|
289
|
|
|
|
|
|
|
=cut |
290
|
|
|
|
|
|
|
|
291
|
|
|
|
|
|
|
|
292
|
|
|
|
|
|
|
sub _run { |
293
|
0
|
|
|
0
|
|
|
my ($self, $fasta_file, $qual_file) = @_; |
294
|
|
|
|
|
|
|
|
295
|
|
|
|
|
|
|
# fasta_file: [midinfo@]sffile|[midinfo@]projectdir|fastafile |
296
|
|
|
|
|
|
|
# qual_file: not supported by newbler |
297
|
|
|
|
|
|
|
|
298
|
|
|
|
|
|
|
# Specify that we want a single ACE output file containing all contigs |
299
|
0
|
|
|
|
|
|
$self->ace(1); |
300
|
|
|
|
|
|
|
|
301
|
|
|
|
|
|
|
# Setup needed files and filehandles first |
302
|
0
|
|
|
|
|
|
my ($output_fh, $output_file) = $self->_prepare_output_file( ); |
303
|
|
|
|
|
|
|
|
304
|
|
|
|
|
|
|
# Set the output directory based on the the output file name |
305
|
0
|
|
|
|
|
|
my $output_dir = dirname($output_file); |
306
|
0
|
|
|
|
|
|
$self->out_dir($output_dir); |
307
|
|
|
|
|
|
|
|
308
|
|
|
|
|
|
|
# Set a log file |
309
|
0
|
|
|
|
|
|
my $log_file = File::Spec->catfile($output_dir, '454Log.txt'); |
310
|
|
|
|
|
|
|
|
311
|
|
|
|
|
|
|
# Get program executable |
312
|
0
|
|
|
|
|
|
my $exe = $self->executable; |
313
|
|
|
|
|
|
|
|
314
|
|
|
|
|
|
|
# Get command-line options |
315
|
0
|
|
|
|
|
|
my $options = $self->_translate_params(); |
316
|
|
|
|
|
|
|
|
317
|
|
|
|
|
|
|
# Usage: runAssembly [options] (sfffile | [regionlist:]analysisDir | readfastafile)... |
318
|
|
|
|
|
|
|
# where options is: [-o projdir] [-nrm] [-p (sfffile | [regionlist:]analysisDir)]... |
319
|
0
|
|
|
|
|
|
my @program_args = ( $exe, @$options, $fasta_file); |
320
|
0
|
|
|
|
|
|
my @ipc_args = ( \@program_args, '>', $log_file ); |
321
|
|
|
|
|
|
|
|
322
|
|
|
|
|
|
|
# Print command for debugging |
323
|
0
|
0
|
|
|
|
|
if ($self->verbose() >= 0) { |
324
|
0
|
|
|
|
|
|
my $cmd = ''; |
325
|
0
|
|
|
|
|
|
$cmd .= join ( ' ', @program_args ); |
326
|
0
|
|
|
|
|
|
for ( my $i = 1 ; $i < scalar @ipc_args ; $i++ ) { |
327
|
0
|
|
|
|
|
|
my $element = $ipc_args[$i]; |
328
|
0
|
|
|
|
|
|
my $ref = ref($element); |
329
|
0
|
|
|
|
|
|
my $value; |
330
|
0
|
0
|
0
|
|
|
|
if ( $ref && $ref eq 'SCALAR') { |
331
|
0
|
|
|
|
|
|
$value = $$element; |
332
|
|
|
|
|
|
|
} else { |
333
|
0
|
|
|
|
|
|
$value = $element; |
334
|
|
|
|
|
|
|
} |
335
|
0
|
|
|
|
|
|
$cmd .= " $value"; |
336
|
|
|
|
|
|
|
} |
337
|
0
|
|
|
|
|
|
$self->debug( "$exe command = $cmd\n" ); |
338
|
|
|
|
|
|
|
} |
339
|
|
|
|
|
|
|
|
340
|
|
|
|
|
|
|
# Execute command |
341
|
0
|
|
|
|
|
|
eval { |
342
|
0
|
0
|
|
|
|
|
IPC::Run::run(@ipc_args) || die("There was a problem running $exe. The ". |
343
|
|
|
|
|
|
|
"error message is: $!."); |
344
|
|
|
|
|
|
|
}; |
345
|
0
|
0
|
|
|
|
|
if ($@) { |
346
|
0
|
|
|
|
|
|
$self->throw("$exe call crashed: $@"); |
347
|
|
|
|
|
|
|
} |
348
|
|
|
|
|
|
|
|
349
|
|
|
|
|
|
|
# Close filehandles |
350
|
0
|
|
|
|
|
|
close($output_fh); |
351
|
|
|
|
|
|
|
|
352
|
|
|
|
|
|
|
# Result files |
353
|
0
|
|
|
|
|
|
my $ace_file = File::Spec->catfile($output_dir, '454Contigs.ace'); |
354
|
0
|
|
|
|
|
|
my $aln_file = File::Spec->catfile($output_dir, '454AlignmentInfo.tsv'); |
355
|
0
|
|
|
|
|
|
my $all_cont_fasta_file = File::Spec->catfile($output_dir, '454AllContigs.fna'); |
356
|
0
|
|
|
|
|
|
my $all_cont_qual_file = File::Spec->catfile($output_dir, '454AllContigs.qual'); |
357
|
0
|
|
|
|
|
|
my $large_cont_fasta_file = File::Spec->catfile($output_dir, '454LargeContigs.fna'); |
358
|
0
|
|
|
|
|
|
my $large_cont_qual_file = File::Spec->catfile($output_dir, '454LargeContigs.qual'); |
359
|
0
|
|
|
|
|
|
my $metrics_file = File::Spec->catfile($output_dir, '454NewblerMetrics.txt'); |
360
|
0
|
|
|
|
|
|
my $status_file = File::Spec->catfile($output_dir, '454ReadStatus.txt'); |
361
|
0
|
|
|
|
|
|
my $progress_file = File::Spec->catfile($output_dir, '454NewblerProgress.txt'); |
362
|
0
|
|
|
|
|
|
my $trim_file = File::Spec->catfile($output_dir, '454TrimStatus.txt'); |
363
|
0
|
|
|
|
|
|
my $sff_dir = File::Spec->catfile($output_dir, 'sff'); |
364
|
|
|
|
|
|
|
|
365
|
|
|
|
|
|
|
# Remove all files except for the ACE file |
366
|
0
|
|
|
|
|
|
for my $file ($aln_file, $all_cont_fasta_file, $all_cont_qual_file, |
367
|
|
|
|
|
|
|
$large_cont_fasta_file, $large_cont_qual_file, $metrics_file, $status_file, |
368
|
|
|
|
|
|
|
$progress_file, $trim_file, $log_file ) { |
369
|
0
|
|
|
|
|
|
unlink $file; |
370
|
|
|
|
|
|
|
} |
371
|
0
|
|
|
|
|
|
rmtree( $sff_dir ); |
372
|
|
|
|
|
|
|
|
373
|
|
|
|
|
|
|
# Move output file to proper location/name |
374
|
0
|
0
|
|
|
|
|
move ($ace_file, $output_file) or $self->throw("Could not move file ". |
375
|
|
|
|
|
|
|
"'$ace_file' to '$output_file': $!"); |
376
|
|
|
|
|
|
|
|
377
|
0
|
|
|
|
|
|
return $output_file; |
378
|
|
|
|
|
|
|
} |
379
|
|
|
|
|
|
|
|
380
|
|
|
|
|
|
|
1; |