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University System of Maryland

Ih-Chang Hsu

James W Shih

William E Highsmith, Jr

A-Lien Lu-Chang

Detection of probe fragment products of basepair mismatch cleavage indicate the presence and sequence of target DNA. Detection of the target is enhanced by amplification through recycling targets by maintaining an assay temperature between the melting point of the target/probe DNA duplex and that of the target/product complex, in the presence of an amplifier comprising ammonium acetate or an amine derivative (for example, diethylamine, piperidine or ammonium carbonate). Cleavage reduces the size of the duplex, and thus lowering its melting point. The amplifier releases the target from the complex, thereby permitting further catalysis of cleavage and effectively amplifying the signal to be detected.

Method of amplifying DNA and RNA mismatch cleavage products

This invention relates to a method for identifying single base pair mismatches in nucleic acids using mismatch endonucleases and a set of labeled oligonucleotide probes which hybridize to mismatch sequences in the target nucleic acid such that a detectable enzyme-nucleic acid-probe complex forms or labeled cleaved fragments form when a base pair mismatch is present.

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Ellicott City, MD, United States of America

Ih-Chang Hsu