目录

batch_brb

Introduction

batch_brb is a command-line application for automated best reciprocal BLAST and phylogenetic analysis using FastTree. Common ortholog identification methods can have organism availability limitations and difficulty identifying divergent sequences. These issues can be overcome with manual searching but this is a time consuming and tedious process making it unsuitable for moderate to larger scale analyses. batch_brb provides tools to automate the data collection process while maintaining flexibility in hit selection criteria, enabling the analysis of greater numbers of sequences.

batch_brb performs ortholog identification using best reciprocal BLAST. Sequences of interest are searched against a user created database. The top x hits per query per organism are extracted and filtered by y coverage of the query where x and y are specified by the user. Identical hits do not contribute to the hit count. These hits are searched against the organism of the original query sequences. The top x hits per query are filtered by y query coverage as above. Where the hits from the first and reverse BLAST match, the sequences are considered orthologs. batch_brb is designed to enable maximum coverage and requires user analysis of hits for the exclusion of mishits and paralogs.

Documentation

See the batch_brb_manual in the documentation folder for usage instructions

Quick start

batch_brb is available as a Bioconda package and requires Conda and Bioconda. On Linux/Unix-like operating system:

conda create -n batch_brb batch_brb
conda activate batch_brb

Change into the directory where batch_brb should be located and run:

batch_brb_setup

Contact info

For general enquiries please contact ebutterfield@dundee.ac.uk, to report issues please use the GitHub issue tracker.

How to cite batch_brb

If you use this software please cite the protocol publication describing it:

Butterfield, E.R., Abbott, J.C., Field, M.C. (2021). Automated Phylogenetic Analysis Using Best Reciprocal BLAST. In: de Pablos, L.M., Sotillo, J. (eds) Parasite Genomics. Methods in Molecular Biology, vol 2369. Humana, New York, NY.

Please also cite the dependency publications, see documentation for a list of dependencies.

Acknowledgements

This software was created by the Wellcome Centre for Anti-Infectives Research using Wellcome Trust funding.

Thank you to Tim Butterfield, Frederik Drost and Michele Tinti for comments on the code and to Ricardo Canavate del Pino and Ning Zhang for help with testing.

Licence

Copyright (C) 2020 University of Dundee

This program is free software: you can redistribute it and/or modify it under the terms of the GNU General Public License as published by the Free Software Foundation, either version 3 of the License, or (at your option) any later version.

This program is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU General Public License for more details.

You should have received a copy of the GNU General Public License along with this program. If not, see http://www.gnu.org/licenses/.

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