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A method for selective cloning of eukaryotic DNA fragments in Escherichia coli by repeated transformation

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Summary

A procedure is described for the selective cloning in E. coli of restriction endonuclease-generated fragments of eukaryotic DNA linked to a bacterial plasmid replicon. The cloning method involves the initial use of tranformation to amplify an unfractionated heterogeneous population of ligated DNA molecules, and the subsequent purification of plasmid chimeras by repeated cycles of transformation and gradient centrifugation. The procedure has enabled the identification and purification of a rare 6.8x106 dalton EcoRI-generated fragment of X. laevis rDNA that is present in about 0.5% of the rDNA tandem repeat units.

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Authors and Affiliations

  1. Department of Medicine, Stanford University School of Medicine, Stanford, California, USA

    Stanley N. Cohen & Annie C. Y. Chang

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  1. Stanley N. Cohen
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  2. Annie C. Y. Chang
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Communicated by L. Lerman

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Cohen, S.N., Chang, A.C.Y. A method for selective cloning of eukaryotic DNA fragments in Escherichia coli by repeated transformation. Molec. Gen. Genet. 134, 133–141 (1974). https://doi.org/10.1007/BF00268415

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  • DOI: https://doi.org/10.1007/BF00268415

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