Cleavage of nucleic acids
First Claim
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1. A method for detecting a target nucleic acid comprising:
- exposing a composition comprising a purified thermostable FEN-1 endonuclease from an archaebacterial species selected from the group consisting of Pyrococcus furiosus and Methanococcus jannaschii and a purified nucleic acid molecule, wherein said purified nucleic acid molecule is capable of hybridizing to a target nucleic acid to form a FEN-1 endonuclease cleavage structure, to a sample suspected of containing said target nucleic acid;
forming a FEN-1 endonuclease cleavage structure; and
cleaving said FEN-1 endonuclease cleavage structure with said purified thermostable FEN-1 endonuclease, such that said target nucleic acid is detected.
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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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Citations
27 Claims
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1. A method for detecting a target nucleic acid comprising:
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exposing a composition comprising a purified thermostable FEN-1 endonuclease from an archaebacterial species selected from the group consisting of Pyrococcus furiosus and Methanococcus jannaschii and a purified nucleic acid molecule, wherein said purified nucleic acid molecule is capable of hybridizing to a target nucleic acid to form a FEN-1 endonuclease cleavage structure, to a sample suspected of containing said target nucleic acid; forming a FEN-1 endonuclease cleavage structure; and cleaving said FEN-1 endonuclease cleavage structure with said purified thermostable FEN-1 endonuclease, such that said target nucleic acid is detected. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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Specification