Cleavage of nucleic acids
The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5′ nucleases and 3′ exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.
1 - 44 . (canceled)
45 . A composition comprising an isolated FEN-1 endonuclease and an isolated polymerase substantially lacking 5′ nuclease activity.
46 . The composition of claim 45 , wherein said FEN-1 endonuclease and said polymerase are contained in a mixture.
47 . The composition of claim 46 , wherein said mixture comprises a reaction mixture.
48 . The composition of claim 45 , further comprising a nucleic acid molecule.
49 . The composition of claim 48 , wherein said nucleic acid molecule comprises a probe oligonucleotide.
50 . The composition of claim 48 , wherein said nucleic acid molecule is attached to a solid surface.
51 . The composition of claim 45 , further comprising a buffer comprising magnesium.
52 . The composition of claim 45 , further comprising a labeled molecule.
53 . The composition of claim 52 , wherein said labeled molecule comprises a labeled nucleic acid molecule.
54 . The composition of claim 53 , wherein said labeled nucleic acid molecule comprises a fluorescent label.
55 . The composition of claim 45 , further comprising first and second nucleic acid molecules.
56 . The composition of claim 55 , wherein said first and second nucleic acid molecules are both configured to hybridize to a target nucleic acid.
57 . The composition of claim 56 , wherein said first and second nucleic acid molecules are configured to hybridize to a target nucleic acid adjacent to one another.
58 . The composition of claim 45 , wherein said isolated polymerase is from a Thermus species.