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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... Full description

Journal Title: SpringerPlus 2016-04-18, Vol.5 (1), p.477-477
Main Author: Mogni, Virginia Y
Other Authors: Kahan, Mariano A , de Queiroz, Luciano Paganucci , Vesprini, José L , Ortiz, Juan Pablo A , Prado, Darién E
Format: Electronic Article Electronic Article
Language: English
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Publisher: Cham: Springer International Publishing
ID: ISSN: 2193-1801
Link: https://www.ncbi.nlm.nih.gov/pubmed/27217992
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recordid: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4835408
title: Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
format: Article
creator:
  • Mogni, Virginia Y
  • Kahan, Mariano A
  • de Queiroz, Luciano Paganucci
  • Vesprini, José L
  • Ortiz, Juan Pablo A
  • Prado, Darién E
subjects:
  • Anacardiaceae
  • Biomedical and Life Sciences
  • Bioquímica y Biología Molecular
  • Botánica
  • Ciencias Biológicas
  • Ciencias de las Plantas
  • DNA isolation
  • Humanities and Social Sciences
  • Methodology
  • multidisciplinary
  • Phylogeny
  • Schinopsis
  • Science
  • Science (multidisciplinary)
  • Sequencing
ispartof: SpringerPlus, 2016-04-18, Vol.5 (1), p.477-477
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 trn L-F, rps 16 and ndh F 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.
language: eng
source:
identifier: ISSN: 2193-1801
fulltext: no_fulltext
issn:
  • 2193-1801
  • 2193-1801
url: Link


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titleOptimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae
creatorMogni, Virginia Y ; Kahan, Mariano A ; de Queiroz, Luciano Paganucci ; Vesprini, José L ; Ortiz, Juan Pablo A ; Prado, Darién E
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descriptionThe 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 trn L-F, rps 16 and ndh F 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.
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subjectAnacardiaceae ; Biomedical and Life Sciences ; Bioquímica y Biología Molecular ; Botánica ; Ciencias Biológicas ; Ciencias de las Plantas ; DNA isolation ; Humanities and Social Sciences ; Methodology ; multidisciplinary ; Phylogeny ; Schinopsis ; Science ; Science (multidisciplinary) ; Sequencing
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abstractThe 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 trn L-F, rps 16 and ndh F 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.
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