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Targeting in linear DNA duplexes with two complementary probe strands for hybrid stability

Nature Genetics volume 3, pages 365–372 (1993)Cite this article

Abstract

A new in vitro hybridization reaction targets two short complementary RecA protein-coated DNA probes to homologous sequences at any position in a linear duplex DNA molecule. Stable hybrids are obtained after RecA protein removal when both complementary probe strands are present in a four-stranded hybrid, but not when one probe strand is present in a three-stranded hybrid. In four-stranded hybrids with one probe strand biotinylated and the other radiolabelled, the deproteinized hybrids can be isolated and detected by affinity capture on streptavidin-coated magnetic beads. RecA-mediated targeting of complementary biotinylated DNA probe strands allows the affinity capture of 48.5-kilobase duplex λ genomic DNA. These reactions provide a means of isolating any desired duplex gene or chromosomal DNA fragment.

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Authors and Affiliations

  1. Cell and Molecular Biology Laboratory, Life Sciences Division, SRI International, Menlo Park, California, 94025, USA

    Elissa P. Sena & David A. Zarling

  2. Department of Laboratory Medicine, University of California at San Francisco, San Francisco, California, 94143, USA

    David A. Zarling

Authors
  1. Elissa P. Sena
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  2. David A. Zarling
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Sena, E., Zarling, D. Targeting in linear DNA duplexes with two complementary probe strands for hybrid stability. Nat Genet 3, 365–372 (1993). https://doi.org/10.1038/ng0493-365

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