COMPARATIVE ANALYSIS OF GENOMIC DNA EXTRACTION METHODS IN LIMA BEAN (Phaseolus lunatus L.)

Authors

Jéssica Daniele Lustosa da Silva, Universidade Federal do PiauíFollow
Gisele Holanda de Sa, Universidade Federal do Piauí
Rafael da Costa Almeida, Universidade Federal do Piauí
Amanda Camila dos Santos Linhares, Universidade Federal do Piauí
Marcos Emanuel Oliveira Bezerra, Universidade Federal do Piauí
Igor Benício Campêlo, Universidade Federal do Piauí
Marilha Vieira de Brito, Universidade Federal do Piauí
Ângela Celis de Almeida Lopes, Universidade Federal do PiauíFollow
Regina Lucia Ferreira Gomes, Universidade Federal do PiauíFollow

Document Type

Article

Date of this Version

3-2017

Citation

ANNUAL REPORT OF THE BEAN IMPROVEMENT COOPERATIVE, No. 60, March 2017. Published by USDA.

Comments

U.S. government work.

Abstract

INTRODUCTION: The lima bean (Phaseolus lunatus L.) is the second most important legume of the genus Phaseolus (Maquet et al. 1999). In Brazil, this species is mainly cultivated in the Northeast region and presenting great socioeconomic importance for family communities. In that region there are water scarcity and predominate, long periods of drought, what favors the selection of drought tolerant genotypes and adaptation to different environments? The development of research on lima bean Genetics, Genetic Resources and Plant Breeding is extremely important considering its socioeconomic potential and the lack of information for its exploitation. Thus, we aimed to compare five genomic DNA extraction methods and to determine the most efficient protocol for Phaseolus lunatus, with a view to genotyping.

MATERIAL AND METHODS: Five protocols of DNA extraction in lima bean were evaluated: Dellaporta et al. (1983); Doyle and Doyle (1987), modified; Khanuja et al. (1999); Ferreira and Grattapaglia (1996), Romano and Brazilian (1999). Quality and integrity of DNA samples of each protocol were observed through the comparative analysis of the intensity and the standard of the bands obtained by agarose (0.8%) gel electrophoresis. The spectrophotometer NanoDrop 2000™ also quantified the samples, which provides the absorbance ratios 260/280 nm in addition to the quantification of the nucleic acids. The gels were stained with GelRed^TM and visualized with a photo-documenter. The accessions used to compare the protocols were UFPI 1007 and UFPI 804, from the Germoplasma Bank of the Federal University of Piauí.