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. 1977 Apr;130(1):148-53.
doi: 10.1128/jb.130.1.148-153.1977.

Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation

D D Womble, D P Taylor, R H Rownd

Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation

D D Womble et al. J Bacteriol. 1977 Apr.

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.

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References

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    1. J Bacteriol. 1971 Jun;106(3):773-83 - PubMed
    1. Bacteriol Rev. 1972 Sep;36(3):361-405 - PubMed
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