Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation
Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation
复制标题
通过染料浮力密度离心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
作者：
D. Womble;D. P. Taylor;R. Rownd
介绍了一种用溴化乙锭-氯化铯密度梯度离心法从细菌裂解液中高回收率地提取脱氧核糖核酸（DNA）的方法。在链霉蛋白酶消化和剪切裂解物后，对于给定菌株中的特定质粒，获得了基本上100%的染色体DNA回收率和可重现的共价闭合环状（CCC）质粒DNA回收率。该方法可用于比较不同质粒-宿主组合的CCC质粒/染色体比值。
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.