Title:	Protocol adaptation for brazil nut (Bertholletia excelsa Bonpl) DNA extraction.
Authors:	GIUSTINA, L. D. CABRAL, J. C. BALDONI, A. B. NEVES, L. G.
Affiliation:	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.
Date Issued:	2018
Citation:	International Journal of Current Research, v. 10, n. 02, p. 65270-65275, 2018.
Description:	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.
Thesagro:	Marcador Molecular Bertholletia Excelsa DNA
Keywords:	B-Mercaptoethanol CTAB Molecular analyze Molecular marker SSR Mato Grosso Sinop
ISSN:	0975-833X
Type of Material:	Artigo de periódico
Access:	openAccess
Appears in Collections:	Artigo em periódico indexado (CPAMT)