Birnboim H C, Doly J
Nucleic Acids Res. 1979 Nov 24;7(6):1513-23. doi: 10.1093/nar/7.6.1513.
A procedure for extracting plasmid DNA from bacterial cells is described. The method is simple enough to permit the analysis by gel electrophoresis of 100 or more clones per day yet yields plasmid DNA which is pure enough to be digestible by restriction enzymes. The principle of the method is selective alkaline denaturation of high molecular weight chromosomal DNA while covalently closed circular DNA remains double-stranded. Adequate pH control is accomplished without using a pH meter. Upon neutralization, chromosomal DNA renatures to form an insoluble clot, leaving plasmid DNA in the supernatant. Large and small plasmid DNAs have been extracted by this method.
本文描述了一种从细菌细胞中提取质粒DNA的方法。该方法操作简单，每天可对100个或更多克隆进行凝胶电泳分析，且所提取的质粒DNA纯度足以被限制性内切酶消化。其原理是使高分子量染色体DNA选择性碱变性，而共价闭环DNA保持双链状态。无需使用pH计即可实现充分的pH控制。中和后，染色体DNA复性形成不溶性凝块，质粒DNA则留在上清液中。大小不同的质粒DNA均已通过该方法提取出来。
