An economical large scale procedure to purifyE. coli amplifiable plasmids for DNA sequencing, in vitro transcription and in vitro mutagenesis
An economical large scale procedure to purifyE. coli amplifiable plasmids for DNA sequencing, in vitro transcription and in vitro mutagenesis
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一种经济的大规模纯化方法。
DOI：
10.1007/bf01950053
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
R. E. Cannon
中科院分区：
文献类型：
--
作者：
Guang;R. E. Cannon
描述了一种可重复且经济的程序，用于获得大量且定量的高度纯化的共价闭合环状质粒 DNA。该过程在几个方面与更常用的方法不同。这些是 a) 避免使用 CsCl、溴化乙锭和超速离心机，b) 通过用碱性 SDS 溶液选择性变性染色体 DNA 来富集质粒 DNA，c) 通过用酸-苯酚提取来富集共价闭合环状质粒 DNA，以及 d) 在用 RNase A 消化后通过分子筛色谱法去除小的降解的 RNA 片段。用于DNA测序、体外转录、转化和体外诱变。
A reproducible and economical procedure for obtaining a large and quantitative yield of highly purified covalently closed circular plasmid DNA is described. The procedure departs in several ways from more commonly used methods. These are a) avoidance of the use of CsCl, ethidium bromide and ultracentrifuge, b) enrichment of the plasmid DNA by selective denaturation of chromosomal DNA with an alkaline-SDS solution, c) enrichment of covalently closed circular plasmid DNA by extraction with acid-phenol, and d) removal of small degraded RNA fragements by molecular sieve chromatography after digestion with RNase A. The plasmid DNA prepared by this new procedure is free of contaminants and has been used for DNA sequencing, in vitro transcription, transformation and in vitro mutagenesis.
无需使用 RNase，即可快速、简单地去除质粒 DNA 中的污染 RNA。
影响因子：
2.9
作者：
Norgard,MV
通讯作者：
Norgard,MV