Cleavage of nucleic acids
First Claim
Patent Images
1. A kit for detecting a target nucleic acid, comprising:
- i) a purified thermostable FEN-1 endonuclease; and
ii) an oligonucleotide probe configured to hybridize to a target nucleic acid to form a FEN-1 endonuclease cleavage structure.
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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.
108 Citations
20 Claims
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1. A kit for detecting a target nucleic acid, comprising:
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i) a purified thermostable FEN-1 endonuclease; and ii) an oligonucleotide probe configured to hybridize to a target nucleic acid to form a FEN-1 endonuclease cleavage structure. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A kit for detecting a target sequence, comprising:
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i) a purified thermostable FEN-1 endonuclease; ii) a target nucleic acid, which comprises in 3′
to 5′
order a first region and a second region;iii) a first oligonucleotide that is at least partially complementary to said first region of said target nucleic acid, and iv) a second oligonucleotide comprising a 5′
portion and a 3′
portion, wherein said 3′
portion is complementary to said second region of said target nucleic acid and wherein said 5′
portion is not complementary to said target nucleic acid;wherein said first and second oligonucleotides form an invasive cleavage structure when hybridized to said target nucleic acid. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification