生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 194-198.

• 研究报告 • 上一篇    下一篇

从非变性聚丙烯酰胺凝胶中快速高效回收DNA片段

周颐, 王忆平   

  1. (北京大学生命科学学院,北京 100871)
  • 收稿日期:2012-10-29 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
  • 作者简介:周颐,女,博士研究生,研究方向:生物化学;E-mail:susie1213@pku.edu.cn
  • 基金资助:
    国家自然科学基金项目(30830005),国家杰出青年科学基金项目(39925017)

Rapid and Efficient Recycling DNA Fragments from Non-denaturing Polyacrylamide Gel

Zhou Yi Wang Yiping   

  1. (College of Life Science,Peking University,Beijing 100871)
  • Received:2012-10-29 Revised:2013-05-24 Published:2013-05-24 Online:2013-05-24
  • About author:王忆平,男,教授,博士生导师,研究方向:固氮基因表达调控的生化基础研究;E-mail:wangyp@pku.edu.cn

摘要: 获得特异性的DNA是开展多种分子生物学试验的前提,具有高分辨率的聚丙烯酰胺凝胶电泳是纯化特异性DNA的首选。借助液氮对凝胶的固化作用,介绍了通过研磨破坏聚丙烯酰胺凝胶结构回收纯化DNA的方法。将通过该方法回收纯化的DNA和采用琼脂糖凝胶电泳回收纯化的DNA,经由聚丙烯酰胺凝胶电泳的进一步检测,结果显示,该方法与琼脂糖凝胶电泳回收法相比,所获得DNA的纯度高,特异性好,且效率相当。在提高DNA特异性的同时也兼具了经济高效耗时少等优点,可以广泛应用。

关键词: 非变性聚丙烯酰胺凝胶, 琼脂糖凝胶, 小片段DNA回收纯化, 凝胶阻滞试验

Abstract: Obtaining specific DNA fragments is a prerequisite to carry out varieties of molecular biology experiments and polyacrylamide gel electrophoresis is the first choice to purify specific DNA owing to its high resolution. In this assay, by exploiting the curing effect of liquid nitrogen, an approach applying grinding to break the structure of polyacrylamide gel electrophoresis to extract and purify DNA is introduced. The DNA purified according to the method above and the counterpart based on the purification of agarose gel electrophoresis are subjected to further detection through polyacrylamide gel electrophoresis, with the result manifesting, the DNA purified via the method above has a higher purity and specificity as well as an equal efficiency comparing with the one purified by agarose gel electrophoresis. The method described here improves the specificity of DNA, and simultaneously being economical, efficient and less time-consuming, thus it worth using widely.

Key words: Non-denaturing polyacrylamide gel, Agarose gel, Small DNA fragment recovery and purification, Electrophoretic mobili-ty shift assay