Daikin Industries, Ltd.

Cornelia J Calhoun

David A Zarling

Elissa P Sena

In situ hybridization method

Diagnostic applications of double D-loop formation

A method of identifying the presence of a known target sequence in double-stranded DNA contained in a fixed cellular or subcellular biological structure. By adding a stable, reporter-labeled RecA/single-stranded probe complex to the structure, the target sequence can be effectively labeled by in situ hybridization, allowing the target sequence to be visualized histologically and microscopically or detected by in situ cytometry or cell sorting flow techniques.

The present invention describes the formation of RecA protein catalyzed double-stranded probe:duplex linear target DNA complexes that are stable to deproteinization. The uses of this stable probe:target complex in diagnostic/DNA detection systems in in vitro and in situ DNA hybridization reactions is discussed. The probe:target complexes are also useful for diagnostic application in RecA protein facilitated DNA amplification reactions.

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San Francisco, CA, United States of America

Cornelia J Calhoun