Methods of detecting a target nucleic acid

Abstract

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.