Data/hora: 25/06/2024 00:58:35

Biblioteca(s): | Embrapa Agrossilvipastoril.
Data corrente: | 18/02/2019
Data da última atualização: | 19/02/2019
Autoria: | GIUSTINA, L. D.; CABRAL, J. C.; BALDONI, A. B.; NEVES, L. G.
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.
Título: | Protocol adaptation for brazil nut (Bertholletia excelsa Bonpl) DNA extraction.
Ano de publicação: | 2018
Fonte/Imprenta: | International Journal of Current Research, v. 10, n. 02, p. 65270-65275, 2018.
ISSN: | 0975-833X
Idioma: | Inglês
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 &#946;-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 &#956;L-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 &#946;-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.
Palavras-Chave: | B-Mercaptoethanol; CTAB; Mato Grosso; Molecular analyze; Molecular marker; Sinop; SSR.
Thesagro: | Bertholletia Excelsa; DNA; Marcador Molecular.
Categoria do assunto: | G Melhoramento Genético
URL: | http://ainfo.cnptia.embrapa.br/digital/bitstream/item/192934/1/2018-cpamt-aisy-baldoni-protocol-adaptation-brazil-bertholletia-dna-extraction.pdf
Registro original: | Embrapa Agrossilvipastoril (CPAMT)
Exemplares: | CPAMT - (1302-1) - UPC - AP - DD -  -  -  - DIS