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Registro Completo |
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Biblioteca(s): |
Embrapa Agrossilvipastoril. |
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Data corrente: |
18/02/2019 |
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Data da última atualização: |
19/02/2019 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
GIUSTINA, L. D.; CABRAL, J. C.; BALDONI, A. B.; NEVES, L. G. |
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Afiliação: |
LUANA DELLA GIUSTINA, ESTUDANTE DE PhD, UFMT, CUIABA; JULIANE COSTA CABRAL, MESTRANDA DA UNEMAT, ALTA FLORESTA; AISY BOTEGA BALDONI TARDIN, CPAMT; LEONARDA GRILLO NEVES, UNEMAT, CACERES, MT. |
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Título: |
Protocol adaptation for brazil nut (Bertholletia excelsa Bonpl) DNA extraction. |
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Ano de publicação: |
2018 |
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Fonte/Imprenta: |
International Journal of Current Research, v. 10, n. 02, p. 65270-65275, 2018. |
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ISSN: |
0975-833X |
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Idioma: |
Inglês |
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Conteúdo: |
Brazil nut (Bertholletia excelsa Bonpl.) is an Amazonian Forest species, which is acknowledged for the commercialization of its almonds. It is an endangered species, fact that reinforces the need of conducting conservation studies. Molecular markers are tools that may be used in conservation studies and the primordial stage preceding the use of molecular markers lies on DNA extraction. A well-designed protocol allows isolating DNA in sufficient amount and quality to conduct molecular studies. Thus, the aim of the current study is to adapt a total DNA extraction protocol to be applied to Brazil nut plants. Tests were conducted at different CTAB (2% and 4%) and β-Mercaptoethanol (0.2%, 0.8% and 1.4%) concentrations; washes using chloroform: isoamyl alcohol (CIA) were performed (one or two washes). The extraction products were confirmed in 0.8% agarose gel and quantified in spectrophotometer (in ng μL-1). Plant tissue (vascular cambium and leaf) was collected from 30 Brazil nut individuals in order to confirm the efficiency of the protocol adapted to the species. Samples were subjected to PCR amplification using 10 microsatellite markers. CTAB and β-Mercaptoethanol at concentrations 4% and 0.2%, respectively, along with one wash using CIA, showed the best results when the quality of the extracted material was assessed through its absorbance ratios. The vascular cambium oxidized more easily than the leaf during the DNA isolation process. Thus, it is essential taking some precautions at the time to handle it during collection and storage to assure process efficiency. MenosBrazil nut (Bertholletia excelsa Bonpl.) is an Amazonian Forest species, which is acknowledged for the commercialization of its almonds. It is an endangered species, fact that reinforces the need of conducting conservation studies. Molecular markers are tools that may be used in conservation studies and the primordial stage preceding the use of molecular markers lies on DNA extraction. A well-designed protocol allows isolating DNA in sufficient amount and quality to conduct molecular studies. Thus, the aim of the current study is to adapt a total DNA extraction protocol to be applied to Brazil nut plants. Tests were conducted at different CTAB (2% and 4%) and β-Mercaptoethanol (0.2%, 0.8% and 1.4%) concentrations; washes using chloroform: isoamyl alcohol (CIA) were performed (one or two washes). The extraction products were confirmed in 0.8% agarose gel and quantified in spectrophotometer (in ng μL-1). Plant tissue (vascular cambium and leaf) was collected from 30 Brazil nut individuals in order to confirm the efficiency of the protocol adapted to the species. Samples were subjected to PCR amplification using 10 microsatellite markers. CTAB and β-Mercaptoethanol at concentrations 4% and 0.2%, respectively, along with one wash using CIA, showed the best results when the quality of the extracted material was assessed through its absorbance ratios. The vascular cambium oxidized more easily than the leaf during the DNA isolation process. Thus, it is essential taki... Mostrar Tudo |
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Palavras-Chave: |
B-Mercaptoethanol; CTAB; Mato Grosso; Molecular analyze; Molecular marker; Sinop; SSR. |
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Thesagro: |
Bertholletia Excelsa; DNA; Marcador Molecular. |
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Categoria do assunto: |
G Melhoramento Genético |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/192934/1/2018-cpamt-aisy-baldoni-protocol-adaptation-brazil-bertholletia-dna-extraction.pdf
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Marc: |
LEADER 02409naa a2200289 a 4500
001 2106148
005 2019-02-19
008 2018 bl uuuu u00u1 u #d
022 $a0975-833X
100 1 $aGIUSTINA, L. D.
245 $aProtocol adaptation for brazil nut (Bertholletia excelsa Bonpl) DNA extraction.$h[electronic resource]
260 $c2018
520 $aBrazil nut (Bertholletia excelsa Bonpl.) is an Amazonian Forest species, which is acknowledged for the commercialization of its almonds. It is an endangered species, fact that reinforces the need of conducting conservation studies. Molecular markers are tools that may be used in conservation studies and the primordial stage preceding the use of molecular markers lies on DNA extraction. A well-designed protocol allows isolating DNA in sufficient amount and quality to conduct molecular studies. Thus, the aim of the current study is to adapt a total DNA extraction protocol to be applied to Brazil nut plants. Tests were conducted at different CTAB (2% and 4%) and β-Mercaptoethanol (0.2%, 0.8% and 1.4%) concentrations; washes using chloroform: isoamyl alcohol (CIA) were performed (one or two washes). The extraction products were confirmed in 0.8% agarose gel and quantified in spectrophotometer (in ng μL-1). Plant tissue (vascular cambium and leaf) was collected from 30 Brazil nut individuals in order to confirm the efficiency of the protocol adapted to the species. Samples were subjected to PCR amplification using 10 microsatellite markers. CTAB and β-Mercaptoethanol at concentrations 4% and 0.2%, respectively, along with one wash using CIA, showed the best results when the quality of the extracted material was assessed through its absorbance ratios. The vascular cambium oxidized more easily than the leaf during the DNA isolation process. Thus, it is essential taking some precautions at the time to handle it during collection and storage to assure process efficiency.
650 $aBertholletia Excelsa
650 $aDNA
650 $aMarcador Molecular
653 $aB-Mercaptoethanol
653 $aCTAB
653 $aMato Grosso
653 $aMolecular analyze
653 $aMolecular marker
653 $aSinop
653 $aSSR
700 1 $aCABRAL, J. C.
700 1 $aBALDONI, A. B.
700 1 $aNEVES, L. G.
773 $tInternational Journal of Current Research$gv. 10, n. 02, p. 65270-65275, 2018.
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Embrapa Agrossilvipastoril (CPAMT) |
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