A Method of Recovering Short DNA Fragments from Non-denaturing Polyacrylamide Gel

doi:10.13560/j.cnki.biotech.bull.1985.2015.05.011

Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (5): 68-72.doi: 10.13560/j.cnki.biotech.bull.1985.2015.05.011

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A Method of Recovering Short DNA Fragments from Non-denaturing Polyacrylamide Gel

Xiang Xiaohua^1，2, Jiao Yushun^1，2, Wu Xinru¹, Cheng Yazeng^1，2, Hou Shuo^1，3, Liu Guanshan¹, Wang Yuanying¹

（1. Tobacco Research Institute of CAAS，Key Laboratory of Tobacco Genetic Improvement and Biotechnology，CAAS，Qingdao 266101；2. Graduate School of Chinese Academy of Agricultural Sciences，Beijing 100081；3. College of Agriculture and Plant Protection，Qingdao Agricultural University，Qingdao 266109）

Received:2014-08-27 Online:2015-05-18 Published:2015-05-18

Abstract

Abstract: The polyacrylamide gel electrophoresis with high resolution is a top choice for isolating and purifying specific short DNA fragments, and also serves as a prerequisite for a variety of subsequent molecular biology experiments. In this study, three methods, i.e., modified boiling method（grinding with liquid nitrogen + boiling + concentrating at low temperature）, traditional boiling method（in which DNA is precipitated with absolute ethanol and 3 mol/L sodium acetate together）, and direct recovery method were used to extract short DNA fragments. The short DNA fragments were used as the template for a second PCR reaction. The results showed that recovered DNA fragments by the modified boiling method had higher purity and finer specificity, and the recovery rate was nearly the same as the traditional boiling method. Replacing the method of precipitation with ethanol and sodium acetate by method of concentrating at low temperature is feasible and novel.

Key words: non-denaturing polyacrylamide gel, short DNA fragments, secondary PCR, modified boiling method, gel recovery

Xiang Xiaohua, Jiao Yushun, Wu Xinru, Cheng Yazeng, Hou Shuo, Liu Guanshan, Wang Yuanying. A Method of Recovering Short DNA Fragments from Non-denaturing Polyacrylamide Gel[J]. Biotechnology Bulletin, 2015, 31(5): 68-72.

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A Method of Recovering Short DNA Fragments from Non-denaturing Polyacrylamide Gel

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