DNA polymerase fusions and uses thereof
View Patent ↗The present invention discloses methods of using DNA polymerase fusions at high pH in PCR, DNA sequencing and mutagenesis protocols.
1. A kit for performing at high pH a method selected from the group consisting of: DNA synthesis; cloning of a DNA synthesis product; sequencing DNA; RT-PCR; and linear or exponential PCR amplification, said kit comprising a DNA polymerase fusion, a high pH buffer, and packaging materials therefor, wherein said high pH buffer ranges from pH 9.3 to 12, and wherein said DNA polymerase fusion comprises a wild type Pyrococcus furiosus DNA polymerase fused to a Sulfolobus solfataricus Sso7d protein, wherein said DNA polymerase fusion functions as a DNA polymerase.
2. The kit of claim 1 , further comprising a PCR enhancing factor and/or an additive.
3. The kit of claim 1 , wherein said high pH buffer ranges from pH 9.5 to 12.
4. The kit of claim 1 , further comprising a non-fusion DNA polymers.
5. The kit of claim 4 , wherein said non-fusion DNA polymerase is a Pfu DNA polymerase.
6. The kit of claim 4 , wherein said high pH buffer ranges from pH 9.5 to 12.
7. The kit of claim 6 , wherein said non-fusion DNA polymerase is a Pfu DNA polymerase.
8. A composition comprising a DNA polymerase fusion and a high pH buffer, wherein said high pH buffer has a pH ranging from 9.3 to 12, and wherein said DNA polymerase fusion comprises a wild type Pyrococcus furiosus DNA polymerase fused to a Sulfolobus solfataricus Sso7d protein, wherein said DNA polymersase fusion functions as a DNA polymers.
9. The composition of claim 8 , wherein said high pH buffer ranges from pH 9.5 to 12.
10. The composition of claim 8 , further comprising a PCR enhancing factor ad/or a additive.
11. The composition of claim 10 , wherein said high pH buffer ranges from pH 9.5 to 12.
12. The composition of claim 8 , further comprising a non-fusion DNA polymerase.
13. The composition of claim 12 , wherein said non-fusion DNA polymerase is a Pfu DNA polymerase.
14. The composition of claim 12 , wherein said high pH buffer ranges from pH 9.5 to 12.
15. The composition of claim 14 , wherein said non-fusion DNA polymerase is a Pfu DNA polymerase.