Methods of detecting a target nucleic acid
The invention relates to compositions and methods of generating a signal indicative of the presence of a target nucleic acid in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid with a probe. The invention also includes the steps of cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid in a sample.
1 - 48 . (canceled)
49 . A composition comprising an isolated FEN-1 endonuclease and an isolated polymerase substantially lacking 5′ nuclease activity.
50 . The composition of claim 49 , wherein said FEN-1 endonuclease and said polymerase are contained in a mixture.
51 . The composition of claim 50 , wherein said mixture comprises a reaction mixture.
52 . The composition of claim 49 , further comprising a nucleic acid molecule.
53 . The composition of claim 52 , wherein said nucleic acid molecule comprises a probe oligonucleotide.
54 . The composition of claim 52 , wherein said nucleic acid molecule is attached to a solid surface.
55 . The composition of claim 49 , further comprising a labeled molecule.
56 . The composition of claim 55 , wherein said labeled molecule comprises a labeled nucleic acid molecule.
57 . The composition of claim 56 , wherein said labeled nucleic acid molecule comprises a fluorescent label.
58 . The composition of claim 49 , further comprising first and second nucleic acid molecules.
59 . The composition of claim 58 , wherein said first and second nucleic acid molecules are both configured to hybridize to a target nucleic acid.
60 . The composition of claim 59 , wherein said first and second nucleic acid molecules are configured to hybridize to a target nucleic acid adjacent to one another.
61 . The composition of claim 49 , wherein said isolated polymerase is from a Thermus species.