通过染料浮力密度离心法从细菌裂解物中获得更准确的共价闭合环状质粒与染色体比率的方法。
Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation.
作者信息
Womble D D, Taylor D P, Rownd R H
出版信息
J Bacteriol. 1977 Apr;130(1):148-53. doi: 10.1128/jb.130.1.148-153.1977.
Abstract
A method that gives high recovery of deoxyribonucleic acid (DNA) from crude bacterial lysates using ethidium bromide-cesium chloride density gradient centrifugation is presented. After Pronase digestion and shearing of the lysate, essentially 100% recovery of chromosomal DNA and a reproducible recovery of covalently closed circular (CCC) plasmid DNA is obtained for a specific plasmid in a given strain. This method should be useful for comparing the CCC plasmid/chromosome ratio of various plasmid-host combinations.
摘要
本文介绍了一种使用溴化乙锭-氯化铯密度梯度离心法从粗制细菌裂解物中高效回收脱氧核糖核酸（DNA）的方法。在蛋白酶K消化和裂解物剪切之后，对于给定菌株中的特定质粒，可实现染色体DNA基本100%的回收率以及共价闭合环状（CCC）质粒DNA可重复的回收率。该方法对于比较各种质粒-宿主组合的CCC质粒/染色体比率应是有用的。
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