Method for detection of a target nucleic acid with a FEN nuclease

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 for detecting a target nucleic acid, comprising: a) a purified thermostable FEN-1 endonuclease; and b) an oligonucleotide probe, said oligonucleotide probe configured to participate in a FEN-1 endonuclease cleavage structure in the presence of said target nucleic acid molecule.  
     
     
         50 . The composition of  claim 49 , wherein said oligonucleotide probe comprises a label.  
     
     
         51 . The composition of  claim 50 , wherein said label is configured to be detectable by a method comprising detection of fluorescence.  
     
     
         52 . The composition of  claim 50 , wherein said label is configured to be detectable by a method comprising detection of mass.  
     
     
         53 . The composition of  claim 50 , wherein said label is configured to be detectable by a method comprising detection of fluorescence energy transfer.  
     
     
         54 . The composition of  claim 50 , wherein said label is configured to be detectable by a method comprising detection selected from the group consisting of detection of radioactivity, luminescence, phosphorescence, fluorescence polarization, and charge.  
     
     
         55 . The composition of  claim 49 , wherein said thermostable FEN-1 endonuclease is from an archaebacterium.  
     
     
         56 . The composition of  claim 55 , wherein said archaebacterium is  Pyrococcus furiosus.    
     
     
         57 . The composition of  claim 55 , wherein said archaebacterium is  Methanococcus jannaschii.    
     
     
         58 . The composition of  claim 49 , further comprising a nucleic acid molecule configured to hybridize to said target nucleic acid upstream of said oligonucleotide probe.  
     
     
         59 . The composition of  claim 49 , wherein said oligonucleotide probe is attached to a solid support.  
     
     
         60 . The composition of  claim 58 , wherein said nucleic acid molecule is attached to a solid support.  
     
     
         61 . The composition of  claim 49 , further comprising a polymerase enzyme.  
     
     
         62 . The composition of  claim 49 , further comprising said target nucleic acid.  
     
     
         63 . The composition of  claim 62 , wherein said target nucleic acid comprises DNA.  
     
     
         64 . The composition of  claim 62 , wherein said target nucleic acid comprises synthetic nucleic acid.  
     
     
         65 . The composition of  claim 64 , wherein said synthetic nucleic acid comprises an amplified nucleic acid.  
     
     
         66 . The composition of  claim 65 , wherein said amplified nucleic acid is produced using a polymerase chain reaction.