一种大规模纯化质粒DNA的简单方法。

A simple procedure for large-scale purification of plasmid DNA.

作者信息

Gómez-Márquez J, Freire M, Segade F

机构信息

Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Santiago, Santiago de Compostela, Spain.

出版信息

Gene. 1987;54(2-3):255-9. doi: 10.1016/0378-1119(87)90494-x.

DOI:10.1016/0378-1119(87)90494-x

PMID:3308638

Abstract

We report a simple, rapid and reliable procedure for large-scale purification of plasmid DNA from non-amplified bacterial cultures. It is a modification of the boiling method of Holmes and Quigley [Anal. Biochem. 114 (1981) 193-197] and involves gel-filtration chromatography using Sephacryl S-1000 for final purification of plasmid DNA. This method does not require CsCl gradients and the recovered plasmids are free of RNA and chromosomal DNA, are supercoiled, retain their biological activity, and are suitable for restriction analysis.

摘要

我们报道了一种从未扩增的细菌培养物中大规模纯化质粒DNA的简单、快速且可靠的方法。它是对Holmes和Quigley的煮沸法[《分析生物化学》114 (1981) 193 - 197]的改进，并且包括使用Sephacryl S - 1000进行凝胶过滤色谱以最终纯化质粒DNA。该方法不需要氯化铯梯度，回收的质粒不含RNA和染色体DNA，是超螺旋的，保留其生物活性，并且适用于限制性分析。