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swga: a primer design toolkit for selective whole genome amplification

MOTIVATION: Population genomic analyses are often hindered by difficulties in obtaining sufficient numbers of genomes for analysis by DNA sequencing. Selective whole-genome amplification (SWGA) provides an efficient approach to amplify microbial genomes from complex backgrounds for sequence acquisit...

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Autores principales: Clarke, Erik L, Sundararaman, Sesh A, Seifert, Stephanie N, Bushman, Frederic D, Hahn, Beatrice H, Brisson, Dustin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870857/
https://www.ncbi.nlm.nih.gov/pubmed/28334194
http://dx.doi.org/10.1093/bioinformatics/btx118
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author Clarke, Erik L
Sundararaman, Sesh A
Seifert, Stephanie N
Bushman, Frederic D
Hahn, Beatrice H
Brisson, Dustin
author_facet Clarke, Erik L
Sundararaman, Sesh A
Seifert, Stephanie N
Bushman, Frederic D
Hahn, Beatrice H
Brisson, Dustin
author_sort Clarke, Erik L
collection PubMed
description MOTIVATION: Population genomic analyses are often hindered by difficulties in obtaining sufficient numbers of genomes for analysis by DNA sequencing. Selective whole-genome amplification (SWGA) provides an efficient approach to amplify microbial genomes from complex backgrounds for sequence acquisition. However, the process of designing sets of primers for this method has many degrees of freedom and would benefit from an automated process to evaluate the vast number of potential primer sets. RESULTS: Here, we present swga, a program that identifies primer sets for SWGA and evaluates them for efficiency and selectivity. We used swga to design and test primer sets for the selective amplification of Wolbachia pipientis genomic DNA from infected Drosophila melanogaster and Mycobacterium tuberculosis from human blood. We identify primer sets that successfully amplify each against their backgrounds and describe a general method for using swga for arbitrary targets. In addition, we describe characteristics of primer sets that correlate with successful amplification, and present guidelines for implementation of SWGA to detect new targets. AVAILABILITY AND IMPLEMENTATION: Source code and documentation are freely available on https://www.github.com/eclarke/swga. The program is implemented in Python and C and licensed under the GNU Public License. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
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spelling pubmed-58708572018-03-29 swga: a primer design toolkit for selective whole genome amplification Clarke, Erik L Sundararaman, Sesh A Seifert, Stephanie N Bushman, Frederic D Hahn, Beatrice H Brisson, Dustin Bioinformatics Original Papers MOTIVATION: Population genomic analyses are often hindered by difficulties in obtaining sufficient numbers of genomes for analysis by DNA sequencing. Selective whole-genome amplification (SWGA) provides an efficient approach to amplify microbial genomes from complex backgrounds for sequence acquisition. However, the process of designing sets of primers for this method has many degrees of freedom and would benefit from an automated process to evaluate the vast number of potential primer sets. RESULTS: Here, we present swga, a program that identifies primer sets for SWGA and evaluates them for efficiency and selectivity. We used swga to design and test primer sets for the selective amplification of Wolbachia pipientis genomic DNA from infected Drosophila melanogaster and Mycobacterium tuberculosis from human blood. We identify primer sets that successfully amplify each against their backgrounds and describe a general method for using swga for arbitrary targets. In addition, we describe characteristics of primer sets that correlate with successful amplification, and present guidelines for implementation of SWGA to detect new targets. AVAILABILITY AND IMPLEMENTATION: Source code and documentation are freely available on https://www.github.com/eclarke/swga. The program is implemented in Python and C and licensed under the GNU Public License. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. Oxford University Press 2017-07-15 2017-02-27 /pmc/articles/PMC5870857/ /pubmed/28334194 http://dx.doi.org/10.1093/bioinformatics/btx118 Text en © The Author 2017. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Original Papers
Clarke, Erik L
Sundararaman, Sesh A
Seifert, Stephanie N
Bushman, Frederic D
Hahn, Beatrice H
Brisson, Dustin
swga: a primer design toolkit for selective whole genome amplification
title swga: a primer design toolkit for selective whole genome amplification
title_full swga: a primer design toolkit for selective whole genome amplification
title_fullStr swga: a primer design toolkit for selective whole genome amplification
title_full_unstemmed swga: a primer design toolkit for selective whole genome amplification
title_short swga: a primer design toolkit for selective whole genome amplification
title_sort swga: a primer design toolkit for selective whole genome amplification
topic Original Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870857/
https://www.ncbi.nlm.nih.gov/pubmed/28334194
http://dx.doi.org/10.1093/bioinformatics/btx118
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