FEN-1 endonucleases, mixtures and cleavage methods
First Claim
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1. A method for treating nucleic acid, comprising:
- a) providing;
i) a purified archaeal FEN-1 endonuclease; and
ii) a nucleic acid substrate;
b) treating said nucleic acid substrate under conditions such that said substrate forms one or more cleavage structures; and
c) reacting said endonuclease with said cleavage structures so that one or more cleavage products are produced,wherein said archaeal FEN-1 endonuclease comprises an Archaeoglobus fulgidus FEN-1 endonuclease.
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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 improved cleavage means for the detection and characterization of nucleic acid sequences. Structure-specific nucleases derived from a variety of thermostable organisms are provided. These structure-specific nucleases are used to cleave target-dependent cleavage structures, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
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Citations
20 Claims
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1. A method for treating nucleic acid, comprising:
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a) providing; i) a purified archaeal FEN-1 endonuclease; and ii) a nucleic acid substrate; b) treating said nucleic acid substrate under conditions such that said substrate forms one or more cleavage structures; and c) reacting said endonuclease with said cleavage structures so that one or more cleavage products are produced, wherein said archaeal FEN-1 endonuclease comprises an Archaeoglobus fulgidus FEN-1 endonuclease. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method for treating nucleic acid, comprising:
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a) providing; i) a purified archaeal FEN-1 endonuclease; ii) a source of a target nucleic acid, said target nucleic acid comprising first and second portions, wherein said second portion is downstream of and contiguous to said first portion; iii) a first oligonucleotide comprising a 5′
portion complementary to said first portion of said target nucleic acid; andiv) a second oligonucleotide comprising a 3′
portion and a 5′
portion, wherein said 5′
portion is complementary to said second portion of said target nucleic acid;b) generating a cleavage structure wherein at least said 5′
portion of said first oligonucleotide is annealed to said first portion of said target nucleic acid and wherein at least said 5′
portion of said second oligonucleotide is annealed to said second portion of said target nucleic acid; andc) exposing said cleavage structure to said FEN-1 endonuclease to generate a cleavage product, wherein said archaeal FEN-1 endonuclease comprises Archaeoglobus fulgidus FEN-1 endonuclease. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20)
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Specification