FEN-1 endonucleases, mixtures and cleavage methods
First Claim
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1. A composition comprising an isolated thermostable structure-specific nuclease having an amino acid sequence consisting of SEQ ID NO:
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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
47 Claims
- 1. A composition comprising an isolated thermostable structure-specific nuclease having an amino acid sequence consisting of SEQ ID NO:
- 3. A composition comprising a purified FEN-1 endonuclease comprising an Archaeoglobus fulgidus FEN-1 endonuclease.
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7. A mixture comprising:
- i) a first structure-specific nuclease, wherein said first nuclease comprises a purified FEN-1 endonuclease; and
ii) a purified second structure-specific nuclease. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
- i) a first structure-specific nuclease, wherein said first nuclease comprises a purified FEN-1 endonuclease; and
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20. A nucleic acid treatment kit, comprising:
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a) a purified Archaeoglobus fulgidus FEN-1 endonuclease; and
b) a solution containing manganese. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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33. A nucleic acid treatment kit comprising:
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a) a composition comprising purified FEN-1 endonuclease;
b) a first oligonucleotide comprising a 5′
portion complementary to a first portion of a target nucleic acid; and
c) a second oligonucleotide comprising;
a 5′
portion complementary to a second portion of said target nucleic acid downstream of and contiguous to said first portion; and
a 3′
portion.- View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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Specification