Invasive cleavage of nucleic acids
First Claim
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1. A method of detecting a target nucleic acid, comprising:
- a) exposing a sample comprising a target nucleic acid to a purified thermostable FEN-1 endonuclease from an archaebacterial species selected from the group consisting of Pyrococcus furiosus and Methanococcus jannaschii to generate a cleavage product, wherein said cleavage product is produced from a probe oligonucleotide hybridized to said target nucleic acid to form a cleavage structure comprising a duplex and a flap; and
b) detecting said cleavage product, wherein said cleavage product is indicative of the presence of said target nucleic acid.
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Abstract
The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
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21 Claims
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1. A method of detecting a target nucleic acid, comprising:
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a) exposing a sample comprising a target nucleic acid to a purified thermostable FEN-1 endonuclease from an archaebacterial species selected from the group consisting of Pyrococcus furiosus and Methanococcus jannaschii to generate a cleavage product, wherein said cleavage product is produced from a probe oligonucleotide hybridized to said target nucleic acid to form a cleavage structure comprising a duplex and a flap; and b) detecting said cleavage product, wherein said cleavage product is indicative of the presence of said target nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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Specification