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US 6,673,616 B1Granted
Methods and compositions for characterizing nucleic acids
AssigneeThird Wave Technologies, Inc.
InventorsJames E. Dahlberg, Mary Ann D. Brow, Victor I. Lyamichev
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Abstract
The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5′ nucleases and 3′ exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.
Claims20
A method for cleaving a complex comprising a nucleic acid hairpin structure within the hairpin structure, comprising:
- a) Providing:
- i) a first complex comprising a first nucleic acid and a second nucleic acid, said first nucleic acid having a first portion, wherein at least said first portion of said first nucleic acid is hybridized to said second nucleic acid;
- ii) a third nucleic acid comprising a hairpin structure;
- and
- iii) a thermostable enzyme comprising 5′ nuclease activity:
- b) cleaving said first complex with the thermostable enzame to generate a first cleavage product;
- c) hybridizing at least a portion of said first cleavage product to at least a portion of said third nucleic acid to form a second complex;
- and
- d) cleaving said second complex within said third nucleic acid hairpin structure.
6 more elements
Dependencies are inferred from the claim text.