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Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae

The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites—i.e. tannins and oleoresins—that ar...

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Autores principales: Mogni, Virginia Y., Kahan, Mariano A., de Queiroz, Luciano Paganucci, Vesprini, José L., Ortiz, Juan Pablo A., Prado, Darién E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835408/
https://www.ncbi.nlm.nih.gov/pubmed/27217992
http://dx.doi.org/10.1186/s40064-016-2118-4
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author Mogni, Virginia Y.
Kahan, Mariano A.
de Queiroz, Luciano Paganucci
Vesprini, José L.
Ortiz, Juan Pablo A.
Prado, Darién E.
author_facet Mogni, Virginia Y.
Kahan, Mariano A.
de Queiroz, Luciano Paganucci
Vesprini, José L.
Ortiz, Juan Pablo A.
Prado, Darién E.
author_sort Mogni, Virginia Y.
collection PubMed
description The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites—i.e. tannins and oleoresins—that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS–ETS fragments. The modifications proposed allowed the extraction of 70–120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-2118-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-48354082016-05-23 Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae Mogni, Virginia Y. Kahan, Mariano A. de Queiroz, Luciano Paganucci Vesprini, José L. Ortiz, Juan Pablo A. Prado, Darién E. Springerplus Methodology The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites—i.e. tannins and oleoresins—that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS–ETS fragments. The modifications proposed allowed the extraction of 70–120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-2118-4) contains supplementary material, which is available to authorized users. Springer International Publishing 2016-04-18 /pmc/articles/PMC4835408/ /pubmed/27217992 http://dx.doi.org/10.1186/s40064-016-2118-4 Text en © Mogni et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Methodology
Mogni, Virginia Y.
Kahan, Mariano A.
de Queiroz, Luciano Paganucci
Vesprini, José L.
Ortiz, Juan Pablo A.
Prado, Darién E.
Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title_full Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title_fullStr Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title_full_unstemmed Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title_short Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
title_sort optimization of dna extraction and pcr protocols for phylogenetic analysis in schinopsis spp. and related anacardiaceae
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835408/
https://www.ncbi.nlm.nih.gov/pubmed/27217992
http://dx.doi.org/10.1186/s40064-016-2118-4
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