Rapid step-gradient purification of mitochondrial DNA
- 1 January 1988
- journal article
- research article
- Published by Springer Nature in Molecular Biology Reports
- Vol. 13 (2) , 117-120
- https://doi.org/10.1007/bf00539059
Abstract
A convenient modification of the step gradient (CsCl/ethidium bomide) procedure is described. This rapid method allows isolation of covalently closed circular DNA separated from contaminating proteins, RNA and chromosomal DNA in ca. 5 h. Large scale preparations can be performed for circular DNA from eukaryotic organelles (mitochondria). The protocol uses organelle pelleting/NaCl-sarcosyl incubation steps for mitochondria followed by a CsCl step gradient and exhibits yields equal to the conventional procedures. It results in DNA sufficiently pure to be used for restriction endonuclease analysis, subcloning, 5′-end labeling, gel retention assays, and various types of hybridization.Keywords
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