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# -*-CPerl-*- |
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# Last changed Time-stamp: <2015-01-08 23:54:55 mtw> |
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package Bio::ViennaNGS; |
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use version; our $VERSION = qv('0.12_09'); |
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1; |
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=head1 NAME |
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Bio::ViennaNGS - A Perl distribution for Next-Generation Sequencing |
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(NGS) data analysis |
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=head1 DESCRIPTION |
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Bio::ViennaNGS is a distribution of Perl modules and utilities for building |
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efficient Next-Generation Sequencing (NGS) analysis pipelines. It covers |
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various aspects of NGS data analysis, including (but not limited to) |
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conversion of sequence annotation, evaluation of mapped data, expression |
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quantification and visualization. |
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The main Bio::ViennaNGS module is shipped with a complementary set of |
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(sub)modules: |
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=over |
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=item L<Bio::ViennaNGS::AnnoC>: A Moose interface for storage and |
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conversion of sequence annotation data. |
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=item L<Bio::ViennaNGS::BamStat>: A L<Moose> based class for |
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collecting mapping statistics. |
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=item L<Bio::ViennaNGS::BamStatSummary>: A L<Moose> interface for |
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processing L<Bio::ViennaNGS::BamStat> objects on a set of BAM files. |
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=item L<Bio::ViennaNGS::Fasta>: Routines for accessing genomic |
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sequences implemented through a L<Moose> interface to |
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L<Bio::DB::Fasta>. |
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=item L<Bio::ViennaNGS::Feature>: A L<Moose> based BED6 wrapper. |
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=item L<Bio::ViennaNGS::FeatureChain>: Yet another L <Moose> class for |
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chaining gene annotation features. |
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=item L<Bio::ViennaNGS::MinimalFeature>: A L<Moose> interface for |
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handling elementary gene annotation. |
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=item L<Bio::ViennaNGS::SpliceJunc>: A collection of routines for |
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alternative splicing analysis. |
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=item L<Bio::ViennaNGS::Tutorial>: A comprehensive tutorial of the |
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L<Bio::ViennaNGS> core routines with real-world NGS data. |
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=item L<Bio::ViennaNGS::UCSC>: Routines for visualization of genomics |
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data with the UCSC genome browser. |
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=item L<Bio::ViennaNGS::Util>: A collection of wrapper routines for |
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commonly used third-party NGS utilities, code for normalization of |
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gene expression values based on read count data and a set of utility |
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functions. |
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=back |
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=head1 UTILITIES |
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L<Bio::ViennaNGS> comes with a collection of command line utilities |
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for accomplishing routine tasks often required in NGS data |
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processing. These utilities serve as reference implementation of the |
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routines implemented throughout the modules and can readily be used |
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for atomic tasks in NGS data processing: |
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=over |
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=item F<assembly_hub_constructor.pl>: The UCSC genome browser offers |
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the possibility to visualize any organism (including organisms that |
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are not included in the standard UCSC browser bundle) through hso |
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called 'Assembly Hubs'. This script constructs Assembly Hubs from |
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genomic sequence and annotation data. |
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=item F<bam_split.pl>: Split (paired-end and single-end) BAM alignment |
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files by strand and compute statistics. Optionally create BED output, |
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as well as normalized bedGraph and bigWig files for coverage |
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visualization in genome browsers (see dependencies on third-patry |
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tools below). |
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=item F<bam_to_bigWig.pl>: Produce bigWig coverage profiles from |
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(aligned) BAM files, explicitly considering strandedness. The most |
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natural use case of this tool is to create strand-aware coverage |
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profiles in bigWig format for genome browser visualization. |
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=item F<bam_uniq.pl>: Extract unique and multi mapping reads from BAM |
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alignment files and create a separate BAM file for both uniqe (.uniq.) |
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and multi (.mult.) mappers. |
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=item F<bed2bedGraph.pl>: Convert BED files to (strand specific) |
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bedGraph files, allowing additional annotation and automatic |
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generation of bedGraph files which can easily be converted to big-type |
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files for easy UCSC visualization. |
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=item F<extend_bed.pl>: Extend genomic features in BED files by a |
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certain number of nucleotides, either on both sides or specifically at |
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the 5' or 3' end, respectively. |
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=item F<gff2bed.pl>: Convert RefSeq GFF3 annotation files to BED12 |
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format. Individual BED12 files are created for each feature type |
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(CDS/tRNA/rRNA/etc.). Tested with RefSeq bacterial GFF3 annotation. |
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=item F<kmer_analysis.pl>: Count k-mers of predefined length in FastQ |
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and Fasta files |
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=item F<MEME_XML_motif_extractor.pl>: Compute simple statistics from |
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MEME XML output and return a list of found motifs with the number of |
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sequences containing those motifs as well as nice ggplot graphs. |
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=item F<motiffinda.pl>: Find motifs in annotated sequence |
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features. The motif can be provided as regular expression. |
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=item F<newUCSCdb.pl>: Create a new genome database (ie. add a novel |
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organism) in a local instance of the UCSC genome browser. |
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=item F<normalize_multicov.pl>: Compute normalized expression data in |
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TPM/RPKM from (raw) read counts in bedtools multicov format. TPM |
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reference: Wagner et al, Theory Biosci. 131(4), pp 281-85 (2012) |
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=item F<sj_visualizer.pl>: Convert splice junctions from mapped |
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RNA-seq data in segemehl BED6 splice junction format to BED12 for easy |
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visualization in genome Browsers. |
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=item F<splice_site_summary.pl>: Identify and characterize splice |
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junctions from RNA-seq data by intersecting them with annotated splice |
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junctions. |
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=item F<newUCSCdb.pl>: Create a new genome database for a locally |
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installed instance of the UCSC genome browser. Based on |
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http://genomewiki.ucsc.edu/index.php/Building_a_new_genome_database |
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=item F<trim_fastq.pl>: Trim sequence and quality string fields in a |
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Fastq file by user defined length. |
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=back |
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=head1 DEPENDENCIES |
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The L<Bio::ViennaNGS> modules and classes depend on a set of Perl |
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modules, some of which are part of the Perl core distribution: |
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=over 16 |
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=item L<Bio::Perl> >= 1.00690001 |
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=item L<Bio::DB::Sam> >= 1.39 |
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=item L<Bio::DB::Fasta> |
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=item L<Bio::Tools::GFF> |
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=item L<File::Basename> |
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=item L<File::Share> |
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=item L<File::Temp> |
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=item L<Path::Class> |
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=item L<IPC::Cmd> |
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=item L<Carp> |
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=item L<Template> |
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=item L<Moose> |
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=item L<Moose::Util::TypeConstraints> |
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=item L<namespace::autoclean> |
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=item L<MooseX::Clone> |
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=item L<Tie::Hash::Indexed> |
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=back |
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In addition the following modules are required by the L<Bio::ViennaNGS> utilities: |
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=over 4 |
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=item L<PerlIO::gzip> |
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=item L<Math::Round> |
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=item L<XML::Simple> |
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=item L<Statistics::R> |
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=back |
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L<Bio::ViennaNGS> uses third-party tools for computing intersections |
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of BED files: F<bedtools intersect> from the |
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L<BEDtools|http://bedtools.readthedocs.org/en/latest/content/tools/intersect.html> |
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suite is used to compute overlaps and F<bedtools sort> is used to sort |
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BED output files. Make sure that those third-party utilities are |
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available on your system, and that hey can be found and executed by |
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the Perl interpreter. We recommend installing the latest version of |
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L<BEDtools|https://github.com/arq5x/bedtools2> on your system. |
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=head1 SOURCE AVAILABILITY |
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Source code for this distribution is available from the L<ViennaNGS |
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Github repository|https://github.com/mtw/Bio-ViennaNGS>. |
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=head1 PAPERS |
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If the L<Bio::ViennaNGS> suite is useful for your work or if you use |
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any component of the distribution in a custom pipeline, please cite |
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the following publication: |
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B<"ViennaNGS - A toolbox for building efficient next-generation sequencing |
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analysis pipelines"> |
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I<Michael T. Wolfinger, Joerg Fallmann, Florian Eggenhofer and Fabian Amman> |
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bioRxiv L<doi:10.1101/013011|http://dx.doi.org/10.1101/013011>. |
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=head1 NOTES |
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The L<Bio::ViennaNGS> suite is actively developed and tested on |
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different flavours of Linux and Mac OS X. We have taken care of |
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writing platform-independent code that should run out of the box on |
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most UNIX-based systems, however we do not have access to machines |
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running Microsoft Windows. As such, we have not tested and will not |
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test Windows compatibility. |
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=head1 SEE ALSO |
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=over 13 |
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=item L<Bio::ViennaNGS::AnnoC> |
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240
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=item L<Bio::ViennaNGS::Bam> |
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=item L<Bio::ViennaNGS::BamStat> |
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=item L<Bio::ViennaNGS::BamStatSummary> |
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=item L<Bio::ViennaNGS::Fasta> |
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=item L<Bio::ViennaNGS::Feature> |
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=item L<Bio::ViennaNGS::FeatureChain> |
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=item L<Bio::ViennaNGS::MinimalFeature> |
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=item L<Bio::ViennaNGS::SpliceJunc> |
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=item L<Bio::ViennaNGS::Util> |
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=item L<Bio::ViennaNGS::Tutorial> |
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=item L<Bio::ViennaNGS::UCSC> |
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=item L<Bio::ViennaNGS::Util> |
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264
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=back |
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=head1 AUTHORS |
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268
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=over |
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=item Michael T. Wolfinger E<lt>michael@wolfinger.euE<gt> |
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=item Jörg Fallmann E<lt>fall@tbi.univie.ac.atE<gt> |
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=item Florian Eggenhofer E<lt>florian.eggenhofer@tbi.univie.ac.atE<gt> |
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276
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=item Fabian Amman E<lt>fabian@tbi.univie.ac.at<gt> |
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278
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=back |
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280
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=head1 COPYRIGHT AND LICENSE |
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281
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282
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Copyright (C) 2014 Michael T. Wolfinger E<lt>michael@wolfinger.euE<gt> |
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284
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This library is free software; you can redistribute it and/or modify |
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it under the same terms as Perl itself, either Perl version 5.12.4 or, |
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at your option, any later version of Perl 5 you may have available. |
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288
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This software is distributed in the hope that it will be useful, but |
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WITHOUT ANY WARRANTY; without even the implied warranty of |
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MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. |
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292
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=cut |
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294
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