A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca”
Autor(a) principal: | |
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Data de Publicação: | 2021 |
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/54133 |
Resumo: | “Sucupira branca” is a plant found in the Brazilian Cerrado and is adapted to low fertility soils, and its fruit extract has anti-inflammatory, healing, antiulcerogenic, antimicrobial, cercaricidal, leishmanicidal and antioxidant activities. Furthermore, it provides protection against oxidative stress, is a natural biocontrol agent of Aedes aegypti, has very resistant wood, is a melliferous plant and has been used in reforestation programs. The development of conservation strategies is important for maintaining diversity in natural populations of “sucupira branca” since these populations are in the process of genetic erosion. Molecular biology techniques, which are important for characterizing the genetic diversity of plants to develop conservation strategies, require sufficient high-quality genomic deoxyribonucleic acid (DNA). This study aimed to compare five methods to extract DNA from “sucupira branca”. The quality and concentration of DNA were revealed by agarose gel electrophoresis, and only the protocols of Dellaporta, Wood and Hicks et al. (1983) and Khanuja et al. (1999) did not result in satisfactory quantities of DNA. When PCR (Polymerase Chain Reaction) was performed with three inter-simple sequence repeat (ISSR) primers, DNA was successfully amplified from extractions performed with the protocols proposed by Doyle and Doyle (1987), Romano and Brasileiro (1998) and Ferreira and Grattapaglia (1995), which are less expensive than commercial purification kits. These protocols resulted in DNA of sufficient quality and quantity after the amplification reactions were performed. |
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A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca”ConservationDNA IsolationExtractionISSR.Agricultural Sciences“Sucupira branca” is a plant found in the Brazilian Cerrado and is adapted to low fertility soils, and its fruit extract has anti-inflammatory, healing, antiulcerogenic, antimicrobial, cercaricidal, leishmanicidal and antioxidant activities. Furthermore, it provides protection against oxidative stress, is a natural biocontrol agent of Aedes aegypti, has very resistant wood, is a melliferous plant and has been used in reforestation programs. The development of conservation strategies is important for maintaining diversity in natural populations of “sucupira branca” since these populations are in the process of genetic erosion. Molecular biology techniques, which are important for characterizing the genetic diversity of plants to develop conservation strategies, require sufficient high-quality genomic deoxyribonucleic acid (DNA). This study aimed to compare five methods to extract DNA from “sucupira branca”. The quality and concentration of DNA were revealed by agarose gel electrophoresis, and only the protocols of Dellaporta, Wood and Hicks et al. (1983) and Khanuja et al. (1999) did not result in satisfactory quantities of DNA. When PCR (Polymerase Chain Reaction) was performed with three inter-simple sequence repeat (ISSR) primers, DNA was successfully amplified from extractions performed with the protocols proposed by Doyle and Doyle (1987), Romano and Brasileiro (1998) and Ferreira and Grattapaglia (1995), which are less expensive than commercial purification kits. These protocols resulted in DNA of sufficient quality and quantity after the amplification reactions were performed.Sucupira branca é encontrada no cerrado brasileiro e é uma planta bem adaptada à solos de baixa fertilidade e o extrato de seus frutos possui atividades antiinflamatória, antiulcerogênica, antimicrobiana, cercaricida, leishmanicida e antioxidante. Além disso, fornece proteção conta estresse oxidativo, é um agente natural de biocontrole do Aedes aegypti, possui madeira muito resistente, é uma planta melífora e tem sido usada em programas de reflorestamento. O desenvolvimento de estratégias de conservação é importante para manter a diversidade em populações naturais de sucupira branca já que estas populações estão sofrendo erosão genética. Técnicas de biologia molecular requerem DNA genômico de alta qualidade e são importantes para caracterizar a diversidade genética de plantas e no desenvolvimento de estratégias de conservação. Este trabalho objetivou comparar cinco métodos de extração de DNA em sucupira branca. A qualidade e concentração do DNA foi revelada por eletroforese em gel de agarose e apenas os protocolos de Dellaporta, Wood and Hicks (1983) e Khanuja et al. (1999) não resultaram em quantidades satisfatórias de DNA. Quando realizou-se a PCR com três primers ISSR, o DNA foi amplificado com sucesso a partir do DNA extraído de acordo com os protocolos propostos por Doyle e Doyle (1987), Ferreira e Grattapaglia (1995) e Romano e Brasileiro (1998), que são menos caros quando comparados a kits comerciais de extração de DNA; fornecendo um DNA de qualidade e em quantidade suficiente para a realização de reações de amplificação de DNA por meio da PCR.EDUFU2021-12-29info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.ufu.br/index.php/biosciencejournal/article/view/5413310.14393/BJ-v37n0a2021-54133Bioscience Journal ; Vol. 37 (2021): Continuous Publication; e37092Bioscience Journal ; v. 37 (2021): Continuous Publication; e370921981-3163reponame:Bioscience journal (Online)instname:Universidade Federal de Uberlândia (UFU)instacron:UFUenghttps://seer.ufu.br/index.php/biosciencejournal/article/view/54133/33016Brazil; Contemporary Copyright (c) 2021 Jaílson do Nascimento Silva, João Paulo Gomes Viana, Marcones Ferreira Costa , Gisele Holanda de Sá, Maria Fernanda da Costa Gomes, Lidiane de Lima Feitoza , Sérgio Emílio dos Santos Valente https://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessdo Nascimento Silva, Jaílson Gomes Viana, João PauloCosta , Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Feitoza , Lidiane de Lima Valente , Sérgio Emílio dos Santos 2022-05-25T13:36:39Zoai:ojs.www.seer.ufu.br:article/54133Revistahttps://seer.ufu.br/index.php/biosciencejournalPUBhttps://seer.ufu.br/index.php/biosciencejournal/oaibiosciencej@ufu.br||1981-31631516-3725opendoar:2022-05-25T13:36:39Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
title |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
spellingShingle |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” do Nascimento Silva, Jaílson Conservation DNA Isolation Extraction ISSR. Agricultural Sciences |
title_short |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
title_full |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
title_fullStr |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
title_full_unstemmed |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
title_sort |
A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca” |
author |
do Nascimento Silva, Jaílson |
author_facet |
do Nascimento Silva, Jaílson Gomes Viana, João Paulo Costa , Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Feitoza , Lidiane de Lima Valente , Sérgio Emílio dos Santos |
author_role |
author |
author2 |
Gomes Viana, João Paulo Costa , Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Feitoza , Lidiane de Lima Valente , Sérgio Emílio dos Santos |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
do Nascimento Silva, Jaílson Gomes Viana, João Paulo Costa , Marcones Ferreira Sá, Gisele Holanda de Gomes, Maria Fernanda da Costa Feitoza , Lidiane de Lima Valente , Sérgio Emílio dos Santos |
dc.subject.por.fl_str_mv |
Conservation DNA Isolation Extraction ISSR. Agricultural Sciences |
topic |
Conservation DNA Isolation Extraction ISSR. Agricultural Sciences |
description |
“Sucupira branca” is a plant found in the Brazilian Cerrado and is adapted to low fertility soils, and its fruit extract has anti-inflammatory, healing, antiulcerogenic, antimicrobial, cercaricidal, leishmanicidal and antioxidant activities. Furthermore, it provides protection against oxidative stress, is a natural biocontrol agent of Aedes aegypti, has very resistant wood, is a melliferous plant and has been used in reforestation programs. The development of conservation strategies is important for maintaining diversity in natural populations of “sucupira branca” since these populations are in the process of genetic erosion. Molecular biology techniques, which are important for characterizing the genetic diversity of plants to develop conservation strategies, require sufficient high-quality genomic deoxyribonucleic acid (DNA). This study aimed to compare five methods to extract DNA from “sucupira branca”. The quality and concentration of DNA were revealed by agarose gel electrophoresis, and only the protocols of Dellaporta, Wood and Hicks et al. (1983) and Khanuja et al. (1999) did not result in satisfactory quantities of DNA. When PCR (Polymerase Chain Reaction) was performed with three inter-simple sequence repeat (ISSR) primers, DNA was successfully amplified from extractions performed with the protocols proposed by Doyle and Doyle (1987), Romano and Brasileiro (1998) and Ferreira and Grattapaglia (1995), which are less expensive than commercial purification kits. These protocols resulted in DNA of sufficient quality and quantity after the amplification reactions were performed. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12-29 |
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/54133 10.14393/BJ-v37n0a2021-54133 |
url |
https://seer.ufu.br/index.php/biosciencejournal/article/view/54133 |
identifier_str_mv |
10.14393/BJ-v37n0a2021-54133 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/54133/33016 |
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 |
EDUFU |
publisher.none.fl_str_mv |
EDUFU |
dc.source.none.fl_str_mv |
Bioscience Journal ; Vol. 37 (2021): Continuous Publication; e37092 Bioscience Journal ; v. 37 (2021): Continuous Publication; e37092 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|| |
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1797069082826113024 |