PDF(Sci-hub)
Abstract：
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.
摘要：
描述了一种从细菌细胞中提取质粒DNA的方法。该方法足够简单，可以通过每天100个或更多克隆的凝胶电泳进行分析，但仍能产生纯度足以被限制酶消化的质粒DNA。该方法的原理是高分子量染色体DNA的选择性碱性变性,而共价闭合的环状DNA保持双链。在不使用pH计的情况下实现充分的pH控制。中和后，染色体DNA再生形成不溶性凝块，在上清液中留下质粒DNA。已通过该方法提取了大的和小的质粒DNA。
