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NAMEBio::Tools::Run::StandAloneBlastPlus - Compute with NCBI's blast+ suite *ALPHA*SYNOPSISNOTE: This module is related to the Bio::Tools::Run::StandAloneBlast system in name (and inspiration) only. You must use this module directly.# existing blastdb: $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'mydb' ); # create blastdb from fasta file and attach $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'mydb', -db_data => 'myseqs.fas', -create => 1 ); # create blastdb from BioPerl sequence collection objects $alnio = Bio::AlignIO->new( -file => 'alignment.msf' ); $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'mydb', -db_data => $alnio, -create => 1 ); @seqs = $alnio->next_aln->each_seq; $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'mydb', -db_data => \@seqs, -create => 1 ); # create database with masks $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'my_masked_db', -db_data => 'myseqs.fas', -masker => 'dustmasker', -mask_data => 'maskseqs.fas', -create => 1 ); # create a mask datafile separately $mask_file = $fac->make_mask( -data => 'maskseqs.fas', -masker => 'dustmasker' ); # query database for metadata $info_hash = $fac->db_info; $num_seq = $fac->db_num_sequences; @mask_metadata = @{ $fac->db_filter_algorithms }; # perform blast methods $result = $fac->tblastn( -query => $seqio ); # see Bio::Tools::Run::StandAloneBlastPlus::BlastMethods # for many more details DESCRIPTIONNOTE: This module requires BLAST+ v. 2.2.24+ and higher. Until the API stabilizes for BLAST+, consider this module highly experimental.This module along with Bio::Tools::Run::StandAloneBlastPlus::BlastMethods allows the user to perform BLAST functions using the external program suite "blast+" (available at <ftp://ftp.ncbi.nlm.nih.gov/blast/executables/blast+/LATEST/>), using BioPerl objects and Bio::SearchIO facilities. This wrapper can prepare BLAST databases as well as run BLAST searches. It can also be used to run "blast+" programs independently. This module encapsulates object construction and production of databases and masks. Blast analysis methods ("blastp, psiblast", etc>) are contained in Bio::Tools::Run::StandAloneBlastPlus::BlastMethods. USAGEThe basic mantra is to (1) create a BlastPlus factory using the "new()" constructor, and (2) perform BLAST analyses by calling the desired BLAST program by name off the factory object. The blast database itself and any masking data are attached to the factory object (step 1). Query sequences and any parameters associated with particular programs are provided to the blast method call (step 2), and are run against the attached database.Factory construction/initializationThe factory needs to be told where the blast+ programs live. The "BLASTPLUSDIR" environment variable will be checked for the default executable directory. The program directory can be set for individual factory instances with the "PROG_DIR" parameter. All the blast+ programs must be accessible from that directory (i.e., as executable files or symlinks).Either the database or BLAST subject data must be specified at object construction. Databases can be pre-existing formatted BLAST dbs, or can be built directly from fasta sequence files or BioPerl sequence object collections of several kinds. The key constructor parameters are "DB_NAME", "DB_DATA", "DB_DIR". To specify a pre-existing BLAST database, use "DB_NAME" alone: $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -DB_NAME => 'mydb' ); The directory can be specified along with the basename, or separately with "DB_DIR": $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -DB_NAME => '~/home/blast/mydb' ); #same as $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -DB_NAME => 'mydb', -DB_DIR => '~/home/blast' ); To create a BLAST database de novo, see "Creating a BLAST database". If you wish to apply pre-existing mask data (i.e., the final ASN1 output from one of the blast+ masker programs), to the database before querying, specify it with "MASK_FILE": $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -DB_NAME => 'mydb', -MASK_FILE => 'mymaskdata.asn' ); Creating a BLAST databaseThere are several options for creating the database de novo using attached data, both before and after factory construction. If a temporary database (one that can be deleted by the "cleanup()" method) is desired, leave out the "-db_name" parameter. If "-db_name" is specified, the database will be preserved with the basename specified.Use "-create =" 1> to create a new database (otherwise the factory will look for an existing database). Use "-overwrite =" 1> to create and overwrite an existing database. Note that the database is not created immediately on factory construction. It will be created if necessary on the first use of a factory BLAST method, or you can force database creation by executing $fac->make_db();
Creating and using mask dataThe blast+ mask utilities "windowmasker", "segmasker", and "dustmasker" are available. Masking can be rolled into database creation, or can be executed later. If your mask data is already created and in ASN1 format, set the "-mask_file" attribute on construction (see "Factory constuction/initialization").To create a mask from raw data or an existing database and apply the mask upon database creation, construct the factory like so: $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'my_masked_db', -db_data => 'myseqs.fas', -masker => 'dustmasker', -mask_data => 'maskseqs.fas', -create => 1 ); The masked database will be created during "make_db". The "-mask_data" parameter can be a FASTA filename or any BioPerl sequence object collection. If the datatype ('nucl' or 'prot') of the mask data is not compatible with the selected masker, an exception will be thrown with a message to that effect. To create a mask ASN1 file that can be used in the "-mask_file" parameter separately from the attached database, use the "make_mask()" method directly: $mask_file = $fac->make_mask(-data => 'maskseqs.fas', -masker => 'dustmasker'); # segmasker can use a blastdb as input $mask_file = $fac->make_mask(-mask_db => 'mydb', -masker => 'segmasker') $fac = Bio::Tools::Run::StandAloneBlastPlus->new( -db_name => 'my_masked_db', -db_data => 'myseqs.fas', -mask_file => $mask_file -create => 1 ); Getting database informationTo get a hash containing useful metadata on an existing database (obtained by running "blastdbcmd -info", use "db_info()":# get info on the attached database.. $info = $fac->db_info; # get info on another database $info = $fac->db_info('~/home/blastdbs/another'); To get a particular info element for the attached database, just call the element name off the factory: $num_seqs = $fac->db_num_sequences; # info on all the masks applied to the db, if any: @masking_info = @{ $fac->db_filter_algorithms }; Accessing the Bio::Tools::Run::BlastPlus factoryThe blast+ programs are actually executed by a Bio::Tools::Run::BlastPlus wrapper instance. This instance is available for peeking and poking in the StandAloneBlastPlus "factory()" attribute. For convenience, "BlastPlus" methods can be run from the "StandAloneBlastPlus" object, and are delegated to the "factory()" attribute. For example, to get the blast+ program to be executed, examine either$fac->factory->command or $fac->command Similarly, the current parameters for the "BlastPlus" factory are @parameters = $fac->get_parameters Cleaning up temp filesTemporary analysis files produced under a single factory instances can be unlinked by running$fac->cleanup; Tempfiles are generally not removed unless this method is explicitly called. "cleanup()" only unlinks "registered" files and databases. All temporary files are automatically registered; in particular, "anonymous" databases (such as $fac->Bio::Tools::Run::StandAloneBlastPlus->new( -db_data => 'myseqs.fas', -create => 1 ); without a "-db_name" specification) are registered for cleanup. Any file or database can be registered with an internal method: $fac->_register_temp_for_cleanup('testdb'); Other Goodies
SEE ALSOBio::Tools::Run::StandAloneBlastPlus::BlastMethods, Bio::Tools::Run::BlastPlusFEEDBACKMailing ListsUser feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to the Bioperl mailing list. Your participation is much appreciated.bioperl-l@bioperl.org - General discussion http://bioperl.org/wiki/Mailing_lists - About the mailing lists SupportPlease direct usage questions or support issues to the mailing list:bioperl-l@bioperl.org 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 BugsReport bugs to the Bioperl bug tracking system to help us keep track of the bugs and their resolution. Bug reports can be submitted via the web:http://redmine.open-bio.org/projects/bioperl/ AUTHOR - Mark A. JensenEmail maj -at- fortinbras -dot- usCONTRIBUTORSAPPENDIXThe rest of the documentation details each of the object methods. Internal methods are usually preceded with a _newTitle : new Usage : my $obj = new Bio::Tools::Run::StandAloneBlastPlus(); Function: Builds a new Bio::Tools::Run::StandAloneBlastPlus object Returns : an instance of Bio::Tools::Run::StandAloneBlastPlus Args : named argument (key => value) pairs: -db : blastdb name db()Title : db Usage : $obj->db($newval) Function: contains the basename of the local blast database Example : Returns : value of db (a scalar string) Args : readonly factory()Title : factory Usage : $obj->factory($newval) Function: attribute containing the Bio::Tools::Run::BlastPlus factory Example : Returns : value of factory (Bio::Tools::Run::BlastPlus object) Args : readonly program_version()Title : program_version Usage : $version = $bedtools_fac->program_version() Function: Returns the program version (if available) Returns : string representing location and version of the program Note : this works around the WrapperBase::version() method conflicting with the -version parameter for SABlast (good argument for not having getter/setters for these) package_version()Title : package_version Usage : $version = $bedtools_fac->package_version() Function: Returns the BLAST+ package version (if available) Returns : string representing BLAST+ package version (may differ from version()) DB methodsmake_db()Title : make_db Usage : Function: create the blast database (if necessary), imposing masking if specified Returns : true on success Args : make_mask()Title : make_mask Usage : Function: create masking data based on specified parameters Returns : mask data filename (scalar string) Args : db_info()Title : db_info Usage : Function: get info for database (via blastdbcmd -info); add factory attributes Returns : hash of database attributes Args : [optional] db name (scalar string) (default: currently attached db) set_db_make_args()Title : set_db_make_args Usage : Function: set the DB make arguments attribute with checking Returns : true on success Args : arrayref or hashref of named arguments set_mask_make_args()Title : set_mask_make_args Usage : Function: set the masker make arguments attribute with checking Returns : true on success Args : arrayref or hasref of named arguments check_db()Title : check_db Usage : Function: determine if database with registered name and dir exists Returns : 1 if db present, 0 if not present, undef if name/dir not set Args : [optional] db name (default is 'registered' name in $self->db) [optional] db directory (default is 'registered' dir in $self->db_dir) no_throw_on_crash()Title : no_throw_on_crash Usage : $fac->no_throw_on_crash($newval) Function: set to prevent an exeception throw on a failed blast program execution Example : Returns : value of no_throw_on_crash (boolean) Args : on set, new value (boolean) Internals_fastize()Title : _fastize Usage : Function: convert a sequence collection to a temporary fasta file (sans gaps) Returns : fasta filename (scalar string) Args : sequence collection _register_temp_for_cleanup()Title : _register_temp_for_cleanup Usage : Function: register a file for cleanup with cleanup() method Returns : true on success Args : a file name or a blastdb basename (scalar string) cleanup()Title : cleanup Usage : Function: unlink files registered for cleanup Returns : true on success Args : AUTOLOADIn this module, "AUTOLOAD()" delegates Bio::Tools::Run::WrapperBase and Bio::Tools::Run::WrapperBase::CommandExts methods (including those of Bio::ParamterBaseI) to the "factory()" attribute:$fac->stderr gives you $fac->factory->stderr If $AUTOLOAD isn't pointing to a WrapperBase method, then AUTOLOAD attempts to return a "db_info" attribute: e.g. $fac->db_num_sequences works by looking in the $fac->db_info() hash. Finally, if $AUTOLOAD is pointing to a blast query method, AUTOLOAD runs "run" with the "-method" parameter appropriately set.
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