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Rapid step-gradient purification of mitochondrial DNA

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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.

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Authors and Affiliations

  1. Institute of Human Genetics, University of the Saar, D-6650, Homburg/Saar, FRG

    Cornelius Welter, Eckart Meese & Nikolaus Blin

Authors
  1. Cornelius Welter
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  2. Eckart Meese
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  3. Nikolaus Blin
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Welter, C., Meese, E. & Blin, N. Rapid step-gradient purification of mitochondrial DNA. Mol Biol Rep 13, 117–120 (1988). https://doi.org/10.1007/BF00539059

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  • DOI: https://doi.org/10.1007/BF00539059

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