一种制备质粒DNA的新的快速方法。

A new rapid procedure for the preparation of plasmid DNA.

作者信息

Edwardson P A, Atkinson T, Lowe C R, Small D A

出版信息

Anal Biochem. 1986 Feb 1;152(2):215-20. doi: 10.1016/0003-2697(86)90400-8.

Abstract

This report describes a simple and efficient procedure for the isolation of plasmid DNA free from chromosomal DNA, cellular RNA, and protein. The technique comprises a modified cleared lysate procedure of D.B. Clewell and D.R. Helinski (1969, Proc. Natl. Acad. Sci. USA, 62, 1159-1166) followed by high-performance liquid chromatography on a Dupont Bioseries GF250 surface stable diol-coated silica gel permeation column (Zorbax) for the final purification of the plasmid DNA. The use of HPLC facilitates rapid and high-resolution separations within 3-4 h. Plasmid DNA produced in this manner retains its biological activity and exhibits yields equal to those obtained by the conventional cesium chloride-ethidium bromide density centrifugation method.

摘要

本报告描述了一种简单有效的从染色体DNA、细胞RNA和蛋白质中分离质粒DNA的方法。该技术包括对D.B. Clewell和D.R. Helinski（1969年，《美国国家科学院院刊》，62卷，1159 - 1166页）改良的清亮裂解液方法，随后在杜邦生物系列GF250表面稳定二醇涂层硅胶渗透柱（Zorbax）上进行高效液相色谱，用于质粒DNA的最终纯化。高效液相色谱的使用有助于在3 - 4小时内实现快速和高分辨率的分离。以这种方式产生的质粒DNA保留其生物活性，并且产量与通过传统的氯化铯 - 溴化乙锭密度离心法获得的产量相当。