Optimized method for dna extraction and PCR amplification in aroeira tree
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Bioscience journal (Online) |
Texto Completo: | https://seer.ufu.br/index.php/biosciencejournal/article/view/62577 |
Resumo: | Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers. |
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Optimized method for dna extraction and PCR amplification in aroeira treeInglêsCTABDNA isolationMyracrodruon urundeuva.Biological SciencesMolecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.Marcadores moleculares são importantes ferramentas na caracterização da diversidade genética de plantas e podem fornecer subsídios para estratégias de conservação de populações ameaçadas de extinção. As diferentes técnicas moleculares envolvem a avaliação de um grande número de indivíduos, portanto, é crucial dispor de métodos rápidos, eficientes e pouco onerosos para a extração de DNA. Dado a importância da aroeira (Myracrodruon urundeuva Fr. All.) faz-se pertinente a otimização de um protocolo que possibilite a obtenção de um DNA íntegro e puro, visando auxiliar estratégias de conservação dessa espécie que se encontra ameaçada de extinção. Dessa forma, o presente trabalho teve como objetivo comparar cinco métodos de extração de DNA: Dellaporta et al. (1983), Doyle e Doyle (1987) modificado, Ferreira e Grattapaglia (1995), Romano e Brasileiro (1999) e Khanuja et al. (1999) e otimizar o protocolo mais eficiente para M. urundeuva. O protocolo de extração de DNA proposto por Doyle e Doyle (1987) modificado, com a utilização de 100 mg de tecido foliar e 6 µl de β-mercaptoetanol foi o protocolo que apresentou as bandas mais nítidas após a eletroforese de DNA e após as reações de amplificação por meio da Polymerase Chain Reaction (PCR). Portanto, sugere-se a utilização do protocolo descrito por Doyle e Doyle (1987) modificado para a extração do DNA de folhas jovens de M. urundeuva para a realização de técnicas que envolvam marcadores moleculares.Universidade Federal de Uberlândia2023-03-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.ufu.br/index.php/biosciencejournal/article/view/6257710.14393/BJ-v39n0a2023-62577Bioscience Journal ; Vol. 39 (2023): Continuous Publication; e39001Bioscience Journal ; v. 39 (2023): Continuous Publication; e390011981-3163reponame:Bioscience journal (Online)instname:Universidade Federal de Uberlândia (UFU)instacron:UFUenghttps://seer.ufu.br/index.php/biosciencejournal/article/view/62577/35649Brazil; Contemporary Copyright (c) 2023 Jailson do Nascimento Silva, Amanda Camila dos Santos Linhares, Dario Abel Palmieri, Marcones Ferreira Costa, Gisele Holanda de Sá, Maria Fernanda da Costa Gomes, Michelli Ferreira dos Santos, Lidiane de Lima Feitoza, Sérgio Emílio dos Santos Valentehttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessSilva, Jailson do NascimentoLinhares, Amanda Camila dos SantosPalmieri, Dario AbelCosta, Marcones FerreiraSá, Gisele Holanda deGomes, Maria Fernanda da CostaSantos, Michelli Ferreira dosFeitoza, Lidiane de Limados Santos Valente, Sérgio Emílio2024-01-31T19:16:18Zoai:ojs.www.seer.ufu.br:article/62577Revistahttps://seer.ufu.br/index.php/biosciencejournalPUBhttps://seer.ufu.br/index.php/biosciencejournal/oaibiosciencej@ufu.br||1981-31631516-3725opendoar:2024-01-31T19:16:18Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Optimized method for dna extraction and PCR amplification in aroeira tree Inglês |
title |
Optimized method for dna extraction and PCR amplification in aroeira tree |
spellingShingle |
Optimized method for dna extraction and PCR amplification in aroeira tree Silva, Jailson do Nascimento CTAB DNA isolation Myracrodruon urundeuva. Biological Sciences |
title_short |
Optimized method for dna extraction and PCR amplification in aroeira tree |
title_full |
Optimized method for dna extraction and PCR amplification in aroeira tree |
title_fullStr |
Optimized method for dna extraction and PCR amplification in aroeira tree |
title_full_unstemmed |
Optimized method for dna extraction and PCR amplification in aroeira tree |
title_sort |
Optimized method for dna extraction and PCR amplification in aroeira tree |
author |
Silva, Jailson do Nascimento |
author_facet |
Silva, Jailson do Nascimento Linhares, Amanda Camila dos Santos Palmieri, Dario Abel Costa, Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Santos, Michelli Ferreira dos Feitoza, Lidiane de Lima dos Santos Valente, Sérgio Emílio |
author_role |
author |
author2 |
Linhares, Amanda Camila dos Santos Palmieri, Dario Abel Costa, Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Santos, Michelli Ferreira dos Feitoza, Lidiane de Lima dos Santos Valente, Sérgio Emílio |
author2_role |
author author author author author author author author |
dc.contributor.author.fl_str_mv |
Silva, Jailson do Nascimento Linhares, Amanda Camila dos Santos Palmieri, Dario Abel Costa, Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Santos, Michelli Ferreira dos Feitoza, Lidiane de Lima dos Santos Valente, Sérgio Emílio |
dc.subject.por.fl_str_mv |
CTAB DNA isolation Myracrodruon urundeuva. Biological Sciences |
topic |
CTAB DNA isolation Myracrodruon urundeuva. Biological Sciences |
description |
Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-03-03 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/62577 10.14393/BJ-v39n0a2023-62577 |
url |
https://seer.ufu.br/index.php/biosciencejournal/article/view/62577 |
identifier_str_mv |
10.14393/BJ-v39n0a2023-62577 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/62577/35649 |
dc.rights.driver.fl_str_mv |
https://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
Brazil; Contemporary |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia |
dc.source.none.fl_str_mv |
Bioscience Journal ; Vol. 39 (2023): Continuous Publication; e39001 Bioscience Journal ; v. 39 (2023): Continuous Publication; e39001 1981-3163 reponame:Bioscience journal (Online) instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
instname_str |
Universidade Federal de Uberlândia (UFU) |
instacron_str |
UFU |
institution |
UFU |
reponame_str |
Bioscience journal (Online) |
collection |
Bioscience journal (Online) |
repository.name.fl_str_mv |
Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU) |
repository.mail.fl_str_mv |
biosciencej@ufu.br|| |
_version_ |
1797069065139781632 |