一种从龙舌兰中分离出的酵母和真菌中提取DNA的快速简便方法。

A rapid and simple method for DNA extraction from yeasts and fungi isolated from Agave fourcroydes.

作者信息

Tapia-Tussell Raul, Lappe Patricia, Ulloa Miguel, Quijano-Ramayo Andrés, Cáceres-Farfán Mirbella, Larqué-Saavedra Alfonso, Perez-Brito Daisy

机构信息

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43 # 130, Col. Chuburná de Hidalgo, Mérida 97200, Yucatán, México.

出版信息

Mol Biotechnol. 2006 May;33(1):67-70. doi: 10.1385/MB:33:1:67.

DOI:10.1385/MB:33:1:67

PMID:16691008

Abstract

A simple and easy protocol for extracting high-quality DNA from different yeast and filamentous fungal species is described. This method involves two important steps: first, the disruption of cell walls by mechanical means and freezing; and second, the extraction, isolation, and precipitation of genomic DNA. The absorbance ratios (A(260)/A(280)) obtained ranged from 1.6 to 2.0. The main objective of this procedure is to extract pure DNA from yeast and filamentous fungi, including those with high contents of proteins, polysaccharides, and other complex compounds in their cell walls. The yield and quality of the DNAs obtained were suitable for micro/minisatellite primer-polymerase chain reaction (MSP-PCR) fingerprinting as well as for the sequence of the D1/D2 domain of the 26S rDNA.

摘要

本文描述了一种从不同酵母和丝状真菌物种中提取高质量DNA的简单易行的方案。该方法包括两个重要步骤：首先，通过机械手段和冷冻破坏细胞壁；其次，提取、分离和沉淀基因组DNA。所获得的吸光度比值（A(260)/A(280)）范围为1.6至2.0。该程序的主要目的是从酵母和丝状真菌中提取纯DNA，包括那些细胞壁中蛋白质、多糖和其他复杂化合物含量高的真菌。所获得的DNA的产量和质量适用于微卫星/小卫星引物聚合酶链反应（MSP-PCR）指纹分析以及26S rDNA的D1/D2结构域的测序。