Compositions and methods for detection of a target nucleic acid sequence
The present invention is directed compositions having a Fen nuclease consisting of a 5′ to 3′ exonuclease and/or endonuclease activity and one or more dNTPs.
1 . A composition comprising an isolated Fen nuclease consisting of a 5′ to 3′ exonuclease and/or endonuclease activity and one or more dNTPs
2 . A composition comprising an isolated Fen nuclease consisting of a 5′ to 3′ exonuclease and/or endonuclease activity and a pyrophosphate.
3 . The composition of claim 1 or 2 , further comprising a labeled oligonucleotide.
4 . The composition of claim 1 or 2 , further comprising a nucleic acid primer.
5 . The composition of claim 3 , further comprising a nucleic acid primer.
6 . The composition of claim 1 or 2 , wherein said fen nuclease is thermostable.
7 . The composition of claim 3 , wherein said fen nuclease is thermostable.
8 . The composition of claim 4 , wherein said fen nuclease is thermostable.
9 . The composition of claim 8 , wherein said fen nuclease is selected from the group consisting of a fen nuclease enzyme of Archaeglobus fulgidus, Methanococcus jannaschii, Pyrococcus furiosus, and Pyrococcus horikoshii.
10 . The composition of claim 1 or 2 , further comprising a nucleic acid polymerase that substantially lacks 5′ to 3′ exonuclease activity.
11 . The composition of claim 10 , wherein said nucleic acid polymerase is thermostable.
12 . The composition of claim 11 , wherein said nucleic acid polymerase is selected from the group consisting of Pfu, Pfu-exo, Taq, Yaq-exo, Deep vent and Deep vent exo.
13 . The composition of claim 4 , wherein said nucleic acid primer is between about 10 to 100 nucleotides in length.
14 . The composition of claim 4 , wherein said nucleic acid primer is between about 17-50 nucleotides in length.
15 . The composition of claim 5 , wherein said nucleic acid primer is between about 17-45 nucleotides in length.
16 . The composition of claim 5 , wherein said nucleic acid primer is between about 10 to 100 nucleotides in length.
17 . The composition of claim 5 , wherein said nucleic acid primer is between about 17-50 nucleotides in length.
18 . The composition of claim 5 , wherein said nucleic acid primer is between about 17-45 nucleotides in length.
19 . The composition of claim 1 , wherein said Fen nuclease is a Fen-1 nuclease.