The subject invention provides novel compositions containing a mixture of
(a) an enzyme that possesses substantial 3' 5' exonuclease activity (b) a
DNA polymerase with less 3' 5' exonuclease activity than the enzyme with
substantial 3' 5' exonuclease activity. Preferably, the DNA polymerase
for inclusion in the compositions are DNA polymerases that substantially
lack 3' 5' exonuclease activity. A preferred embodiment of the invention
is a composition comprising the Taq DNA polymerase (isolated from Thermus
aquaticus) and the Pfu DNA polymerase (isolated from Pyrococcus
furiosus). Another aspect of the invention is to provide methods for
synthesizing polynucleotides, typically DNA, using compositions
comprising an enzyme that possesses substantial 3' 5' exonuclease
activity and a DNA polymerase with less 3' 5' exonuclease activity than
the enzymes possessing substantial 3' 5' exonuclease activity, preferably
a DNA polymerase that substantially lacks 3' 5' exonuclease activity.
Another aspect of the invention involves the use the subject method of
polynucleotide synthesis to carry out the synthesis step in a polymerase
chain reaction experiment. Yet another aspect of the invention is to
provide kits for the synthesis of polynucleotides, wherein the kits
comprise an enzyme that possesses substantial 3' 5' exonuclease activity
and a DNA polymerase with less 3' 5' exonuclease activity than the enzyme
possessing substantial 3' 5' exonuclease activity.