NAME
Bio::Tools::Run::Phrap - a wrapper for running Phrap
SYNOPSIS
use Bio::Tools::Run::Phrap;# Run Phrap using an input FASTA filemy $factory = Bio::Tools::Run::Phrap->new( -penalty => -2, -raw => 1 );my $asm_obj = $factory->run($fasta_file, $qual_file);# An assembly object is returned by defaultfor my $contig ($assembly->all_contigs) {... do something ...}# Read some sequencesuse Bio::SeqIO;my $sio = Bio::SeqIO->new(-file => $fasta_file, -format => 'fasta');my @seqs;while (my $seq = $sio->next_seq()) {push @seqs,$seq;}# Run Phrap using input sequence objects and returning an assembly filemy $asm_file = 'results.phrap';$factory->out_type($asm_file);$factory->run(\@seqs);
DESCRIPTION
Wrapper module for the Phrap assembly programPhrap is available at: http://www.phrap.org/
FEEDBACK
Mailing Lists
User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to one of the Bioperl mailing lists. Your participation is much appreciated.
[email protected] - General discussionhttp://bioperl.org/wiki/Mailing_lists - About the mailing lists
Support
Please direct usage questions or support issues to the mailing list:
rather than to the module maintainer directly. Many experienced and reponsive experts will be able look at the problem and quickly address it. Please include a thorough description of the problem with code and data examples if at all possible.
Reporting Bugs
Report bugs to the Bioperl bug tracking system to help us keep track the bugs and their resolution. Bug reports can be submitted via the web:
http://redmine.open-bio.org/projects/bioperl/
AUTHOR - Shawn Hoon
Email shawnh-at-stanford.edu
APPENDIX
The rest of the documentation details each of the objectmethods. Internal methods are usually preceded with a _
new
Title : newUsage : $factory = Bio::Tools::Run::Phrap->new(-penalty => -2, # parameter option and value-raw => 1 # flag (1=yes, 0=no));Function: Create a new Phrap factoryReturns : A Bio::Tools::Run::Phrap objectArgs : Phrap options available in this module:
Option names & default values taken from the PHRAP manual:
1. Scoring of pairwise alignments
-penalty -2Mismatch (substitution) penalty for SWAT comparisons.-gap_init penalty-2Gap initiation penalty for SWAT comparisons.-gap_ext penalty-1Gap extension penalty for SWAT comparisons.-ins_gap_ext gap_extInsertion gap extension penalty for SWAT comparisons (insertion insubject relative to query).-del_gap_ext gap_extDeletion gap extension penalty for SWAT comparisons (deletion insubject relative to query).-matrix [None]Score matrix for SWAT comparisons (if present, supersedes -penalty)-raw *Use raw rather than complexity-adjusted Smith-Waterman scores.
2. Banded search
-maxmatch 30Maximum length of matching word. For cross_match, the default valueis equal to minmatch, instead of 30.-max_group_size 20Group size (query file, forward strand words)-word_raw *Use raw rather than complexity-adjusted word length, in testingagainst minmatch (N.B. maxmatch always refer to raw lengths). (Thedefault is to adjust word length to reflect complexity of matchingsequence).-bandwidth 141/2 band width for banded SWAT searches (full width is 2 timesbandwidth + 1). Decreasing bandwidth also decreases running time atthe expense of sensitivity. Phrap assemblies of clones containing longtandem repeats of a short repeat unit (< 30 bp) may be more accuratelyassembled by decreasing -bandwidth; -bandwidth should be set such that2 bandwidth + 1 is less than the length of a repeat unit. -bandwidth 0can be used to find gap-free alignments.
3. Filtering of matches
-minscore 30Minimum alignment score.-vector_bound 80Number of potential vector bases at beginning of each read. Matchesthat lie entirely within this region are assumed to represent vectormatches and are ignored. For cross_match, the default value is 0instead of 80.-masklevel 80(cross_match only). A match is reported only if at least (100 -masklevel)% of the bases in its "domain" (the part of the query thatis aligned) are not contained within the domain of any higher-scoringmatch.Special cases:-masklevel 0 report only the single highest scoring match for each query-masklevel 100 report any match whose domain is not completely containedwithin a higher scoring match-masklevel 101 report all matches
4. Input data interpretation
-default_qual 15Quality value to be used for each base, when no input .qual file isprovided. Note that a quality value of 15 corresponds to an error rateof approximately 1 in 30 bases, i.e. relatively accurate sequence. Ifyou are using sequence that is substantially less accurate than thisand do not have phred-generated quality values you should be sure todecrease the value of this parameter.-subclone_delim .(phrap only). Subclone name delimiter: Character used to indicate endof that part of the read name that corresponds to the subclone name-n_delim 1(phrap only). Indicates which occurrence of the subclone delimitercharacter denotes the end of the subclone name (so for example-subclone_delim _ -n_delim 2means that the end of the subclone name occurs at thesecond occurrence of the character '_'). Must be the same for allreads!-group_delim _(phrap only). Group name delimiter: Character used to indicate end ofthat part of the read name that corresponds to the group name(relevant only if option -preassemble is used); this character mustoccur before the subclone delimiter (else it has no effect, and theread is not assigned to a group).-trim_start 0(phrap only). No. of bases to be removed at beginning of each read.
5. Assembly
-forcelevel 0(phrap only). Relaxes stringency to varying degree during finalcontig merge pass. Allowed values are integers from 0 (moststringent) to 10 (least stringent), inclusive.-bypasslevel 1(phrap only). Controls treatment of inconsistent reads in merge.Currently allowed values are 0 (no bypasses allowed; most stringent)and 1 (a single conflicting read may be bypassed).-maxgap 30(phrap only). Maximum permitted size of an unmatched region inmerging contigs, during first (most stringent) merging pass.-repeat_stringency .95(phrap only). Controls stringency of match required for joins. Mustbe less than 1 (highest stringency), and greater than 0 (loweststringency).-revise_greedy *(phrap only). Splits initial greedy assembly into pieces at "weakjoins", and then tries to reattach them to give higher overall score.Use of this option should correct some types of missassembly.-shatter_greedy *(phrap only). Breaks assembly at weak joins (as with -revise_greedy)but does not try to reattach pieces.-preassemble *(phrap only). Preassemble reads within groups, prior to merging withother groups. This is useful for example when the input data setconsists of reads from two distinct but overlapping clones, and it isdesired to assemble the reads from each clone separately beforemerging in order to reduce the risk of incorrect joins due torepeats. The preassemble merging pass is relatively stringent and notguaranteed to merge all of the reads from a group.Groups are indicated by the first part of the read name, up to thecharacter specified by -group_delim.-force_high *(phrap only). Causes edited high-quality discrepancies to be ignoredduring final contig merge pass. This option may be useful when it issuspected that incorrect edits are causing a misassembly.
6. Consensus sequence construction
-node_seg 8(phrap only). Minimum segment size (for purposes of traversingweighted directed graph).-node_space 4(phrap only). Spacing between nodes (in weighted directed graph).
7. Output
Not implemented in this Perl module.
8. Miscellaneous
-retain_duplicates *(phrap only). Retain exact duplicate reads, rather than eliminatingthem.-max_subclone_size 5000(phrap only). Maximum subclone size -- for forward-reverse read pairconsistency checks.-trim_penalty -2(phrap only). Penalty used for identifying degenerate sequence atbeginning & end of read.-trim_score 20(phrap only). Minimum score for identifying degenerate sequence atbeginning & end of read.-trim_qual 13(phrap only). Quality value used in to define the "high-quality" partof a read, (the part which should overlap; this is used to adjustqualities at ends of reads.-confirm_length 8(phrap only). Minimum size of confirming segment (segment starts at3d distinct nuc following discrepancy).-confirm_trim 1(phrap only). Amount by which confirming segments are trimmed atedges.-confirm_penalty -5(phrap only). Penalty used in aligning against "confirming" reads.-confirm_score 30(phrap only). Minimum alignment score for a read to be allowed to"confirm" part of another read.-indexwordsize 10Size of indexing (hashing) words, used in finding word matchesbetween sequences. The value of this parameter has a generally minoreffect on run time and memory usage.
out_type
Title : out_typeUsage : $assembler->out_type('Bio::Assembly::ScaffoldI')Function: Get/set the desired type of outputReturns : The type of results to returnArgs : Desired type of results to return (optional):'Bio::Assembly::IO' object'Bio::Assembly::ScaffoldI' object (default)The name of a file to save the results in
run
Title : runUsage : $asm = $factory->run($fasta_file)Function: Run PhrapReturns : Assembly results (file, IO object or assembly object)Args : - sequence input (FASTA file or sequence object arrayref)- optional quality score input (QUAL file or quality score objectarrayref)
_run
Title : _runUsage : $factory->_run()Function: Make a system call and run PhrapReturns : An assembly fileArgs : - FASTA file- optional QUAL file