DNA polymerases with enhanced length of primer extension
View Patent ↗A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3′-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3′-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.
1. A kit for the synthesis of a polynucleotide, said kit comprising A mixture comprising:
(a) a first DNA polymerase, wherein said first polymerase possesses 3′-5′ exonuclease activity selected from the group consisting of Archaebacterial DNA polymerases, and
(b) a second DNA polymerase, wherein said second polymerase lacks 3′-5′ exonuclease activity selected from the group consisting of thermostable DNA polymerases lacking 3′-5′ exonuclease activity; wherein the ratio of DNA polymerase activity of the first DNA polymerase to the DNA polymerase activity of the second DNA polymerase is from about 1:100 up to about 1:600.
2. A kit according to claim 1 , wherein said Thermus aquaticus DNA polymerase is selected from the group consisting of wild-type Thermus aquaticus DNA polymerase and N-terminal deleted forms of the same enzyme.
3. A kit according to claim 1 , wherein said first DNA polymerase comprises Pyrococcus furiosus DNA polymerase.
4. A kit according to claim 1 , wherein said second DNA polymerase comprises Thermus aquaticus DNA polymerase.
5. A kit according to claim 2 , wherein said Thermus aquaticus DNA polymerase comprises Klentaq-278 DNA polymerase.