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一种从琼脂糖凝胶中回收DNA片段的优化冻挤法。

An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels.

作者信息

Tautz D, Renz M

出版信息

Anal Biochem. 1983 Jul 1;132(1):14-9. doi: 10.1016/0003-2697(83)90419-0.

Abstract

A procedure for quick and simple elution of DNA from agarose gels is presented. After electrophoresis, bands of interest are cut out of the gel and the slices are equilibrated in a neutral salt buffer. The slices are then frozen and centrifuged through a filtration assembly whereby the DNA-containing buffer is squeezed out. The method is simple, quick, and suitable for the safe handling of small amounts of DNA (less than 1 microgram). The isolated DNA is susceptible to any enzymatic reaction and also to chemical sequencing. The method is most useful for rapid preparation of specifically end-labeled DNA fragments (e.g., for sequencing), but may also be utilized for any other preparative applications.

摘要

本文介绍了一种从琼脂糖凝胶中快速简单洗脱DNA的方法。电泳后,从凝胶中切下感兴趣的条带,将切片在中性盐缓冲液中平衡。然后将切片冷冻并通过过滤组件离心,从而挤出含DNA的缓冲液。该方法简单、快速,适用于安全处理少量DNA(少于1微克)。分离出的DNA易于进行任何酶促反应以及化学测序。该方法对于快速制备特异性末端标记的DNA片段(例如用于测序)最为有用,但也可用于任何其他制备应用。

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