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从土壤中提取微生物DNA用于PCR扩增的方法。

Methods for microbial DNA extraction from soil for PCR amplification.

作者信息

Yeates C, Gillings M R, Davison A D, Altavilla N, Veal D A

机构信息

Key Centre for Biodiversity and Bioresources, School of Biological Sciences. Macquarie University, Sydney, NSW 2109. Australia.

出版信息

Biol Proced Online. 1998 May 14;1:40-47. doi: 10.1251/bpo6.

Abstract

Amplification of DNA from soil is often inhibited by co-purified contaminants. A rapid, inexpensive, large-scale DNA extraction method involving minimal purification has been developed that is applicable to various soil types (1). DNA is also suitable for PCR amplification using various DNA targets. DNA was extracted from 100g of soil using direct lysis with glass beads and SDS followed by potassium acetate precipitation, polyethylene glycol precipitation, phenol extraction and isopropanol precipitation. This method was compared to other DNA extraction methods with regard to DNA purity and size.

摘要

土壤中DNA的扩增常常受到共纯化污染物的抑制。现已开发出一种快速、廉价、大规模且纯化步骤极少的DNA提取方法,该方法适用于各种土壤类型(1)。提取的DNA也适用于使用各种DNA靶标的PCR扩增。采用玻璃珠和十二烷基硫酸钠直接裂解,随后进行醋酸钾沉淀、聚乙二醇沉淀、苯酚抽提和异丙醇沉淀的方法,从100克土壤中提取DNA。就DNA纯度和大小而言,将该方法与其他DNA提取方法进行了比较。

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