一种纯化微球菌质粒DNA的经济高效的大规模方法。

An economical large-scale procedure to purify Micrococcus plasmid DNA.

作者信息

Verma V, Qazi G N, Parshad R, Chopra C L

机构信息

Genetic Engineering Unit, Regional Research Laboratory, CSIR, Jammu-Tawi, India.

出版信息

Biotechniques. 1988 Nov-Dec;6(10):936, 938, 940.

PMID:2856199

Abstract

A reproducible and economical procedure for obtaining a large yield of highly purified covalently closed circular (ccc) plasmid DNA from an industrially important strain of Micrococcus is described. The procedure adopted here departs in several ways from commonly used protocols for isolation of plasmid DNA from Gram (positive) and Gram (negative) bacteria. The plasmid DNA prepared by this procedure is free of contaminants and is pure enough to be used for electron microscopy, DNA transformation, sequencing, in vitro transcription and mutagenesis.

摘要

本文描述了一种可重复且经济的方法，用于从一种具有工业重要性的微球菌菌株中大量获得高度纯化的共价闭合环状（ccc）质粒DNA。这里采用的方法在几个方面不同于从革兰氏（阳性）和革兰氏（阴性）细菌中分离质粒DNA的常用方案。通过该方法制备的质粒DNA不含污染物，纯度足以用于电子显微镜检查、DNA转化、测序、体外转录和诱变。