Banca de QUALIFICAÇÃO: LARISSA LEMES DOS SANTOS

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : LARISSA LEMES DOS SANTOS
DATA : 12/12/2023
HORA: 13:30
LOCAL: Alta Floresta, MT.
TÍTULO:

Establishment of a DNA extraction protocol and analysis of the genetic diversity of Mouriri guianensis Aubl.

PALAVRAS-CHAVES:

Nucleic acid, Genetic diversity, Melastomataceae, Roncador.

PÁGINAS: 40
GRANDE ÁREA: Ciências Biológicas
ÁREA: Genética
SUBÁREA: Genética Vegetal
RESUMO:

The DNA extraction method using Cetyl Trimethyl Ammonium Bromide (CTAB) is the most accepted protocol for isolating plant DNA. However, adjustments are necessary to optimize the process, especially in species that do not have an established protocol, such as the roncador (Mouriri guianensis Aubl.). The roncador is a fruitful species and considered a medicinal plant, belonging to the Melastomataceae family and with great economic and ecological potential. Therefore, the objective of the work was to establish a DNA extraction protocol for roncador, aiming for subsequent molecular studies. Genetic analyzes were carried out on native populations of the roncador with natural occurrence in the state of Mato Grosso, Brazil. DNA extraction was based on the CTAB method described by Doyle & Doyle (1990) with the addition of polyvinylpyrrolidone (PVP). To determine the best protocol, changes in the concentration of CTAB (3% and 5%) and the presence and absence of Proteinase K were tested. The concentration of extracted DNA and the A260/A280 and A260/A230 absorbance ratios were estimated by spectrophotometry (ND-3800-OD Nano DOT) and its integrity analyzed by electrophoresis in 1% agarose gel, prepared in 1x TBE buffer (Tris-Borate-EDTA) and stained with ethidium bromide (10 mg mL-1). The concentration of CTAB in the extraction buffer interfered with the quality of DNA extracted from the roncador. The increase in the CTAB concentration to 5% was efficient, and enabled a DNA extraction of greater integrity, with clear and strong bands, and without vertical dragging in the gel, however, it did not influence the amount of DNA obtained by spectrophotometry. Regarding DNA purity, the average of the samples from the tested protocols presented values within the range considered optimal, the A260/A280 ratio (1.8 to 2.0), while for A260/A230 ratios, average values below 1 were obtained. ,8. The addition of Proteinase K showed no difference in the integrity of the extracted DNA, therefore, we suggest that this reagent may or may not be added. The quality of the band pattern presented demonstrates that the extraction methods used were satisfactory, especially with CTAB 5% and can be used in molecular studies of roncador. In fact, it is a process that is underway to complete the research.

MEMBROS DA BANCA:
Presidente - 32143101 - ANA APARECIDA BANDINI ROSSI
Externo ao Programa - 221762013 - JULIANA DE FREITAS ENCINAS DARDENGO
Externo à Instituição - 019.240.091-61 - ELIANE CRISTINA MORENO DE PEDRI - UNIFRAN
Externo à Instituição - JULLIANE DUTRA MEDEIROS - UFJF