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从琼脂糖凝胶中提取核酸——四种不同方法的定量和定性比较

Extraction of nucleic acids from agarose gel--a quantitative and qualitative comparison of four different methods.

作者信息

Pun K K, Kam W

机构信息

Department of Medicine, University of California, San Francisco 94123.

出版信息

Prep Biochem. 1990;20(2):123-35. doi: 10.1080/00327489008050184.

Abstract

Agarose gel electrophoresis is commonly used to separate different species of nucleic acids. We compare four different methods of extraction which are commonly used. These methods include buffer extraction, electroelution, glass bead extraction and extraction of DNA from low-melting agarose. The results show that DNA extracted by these four methods is comparable in their ligability to the PMT 21 vectors and the plasmids with insert can be used for subsequent transfections of competent bacteria. There is a higher yield for buffer extraction and electroelution when compared with glass bead extraction and low melting agarose (p less than 0.05). To conclude, the four commonly used methods for DNA isolation are comparable qualitatively. But the simplest method, namely buffer extraction, has the highest yield.

摘要

琼脂糖凝胶电泳常用于分离不同种类的核酸。我们比较了四种常用的提取方法。这些方法包括缓冲液提取、电洗脱、玻璃珠提取以及从低熔点琼脂糖中提取DNA。结果表明,通过这四种方法提取的DNA在与PMT 21载体连接的能力方面具有可比性,并且带有插入片段的质粒可用于后续感受态细菌的转染。与玻璃珠提取和低熔点琼脂糖提取相比,缓冲液提取和电洗脱的产量更高(p小于0.05)。总之,四种常用的DNA分离方法在质量上具有可比性。但最简单的方法,即缓冲液提取,产量最高。

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