Nucleic acid cleavage assays
First Claim
Patent Images
1. A method for detecting a target nucleic acid, comprising:
- a) generating a cleavage structure that is cleavable by Mja FEN-1 endonuclease, wherein said cleavage structure comprises;
i. said target nucleic acid; and
ii. a displaced 5′
nucleic acid arm;
b) cleaving said cleavage structure with a thermostable 5′
nuclease that lacks synthetic activity to release said displaced 5′
nucleic acid arm; and
c) detecting the cleavage of said cleavage structure to detect said target nucleic acid.
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Abstract
The present invention provides novel cleavage agents and polymerases for the cleavage and modification of nucleic acid. The cleavage agents and polymerases find use, for example, for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. In some embodiments, the 5′ nuclease activity of a variety of enzymes is used to cleave a target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
55 Citations
75 Claims
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1. A method for detecting a target nucleic acid, comprising:
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a) generating a cleavage structure that is cleavable by Mja FEN-1 endonuclease, wherein said cleavage structure comprises;
i. said target nucleic acid; and
ii. a displaced 5′
nucleic acid arm;
b) cleaving said cleavage structure with a thermostable 5′
nuclease that lacks synthetic activity to release said displaced 5′
nucleic acid arm; and
c) detecting the cleavage of said cleavage structure to detect 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, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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41. A method for detecting a target nucleic acid, comprising:
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a) generating a cleavage structure that is cleavable by Mja FEN-1 endonuclease, wherein said cleavage structure comprises;
iii. said target nucleic acid; and
iv. a displaced 5′
nucleic acid arm;
b) cleaving said cleavage structure with a thermostable FEN-1 nuclease to release said displaced 5′
nucleic acid arm; and
c) detecting the cleavage of said cleavage structure to detect said target nucleic acid. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75)
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Specification