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Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase

NA polymerases using relatively short extension times and 25 to 35 PCR cycles.

Fidelity of PfuTurbo DNA Polymerase

The error rates of a number of DNA polymerases have been measured previously in a PCR-based forward mutation assay that utilizes the lacIOlacZa target gene.1 In these studies, Pfu DNA polymerase was found to exhibit an average error rate that was 2-fold lower than that of VentR and Deep VentR DNA polymerases,1 3- to 6-fold lower than those of DNA polymerase mixtures1,4,5,7 and 6-fold lower than that of Taq DNA polymerase.1

The effect of the PCR enhancing factor on the replication fidelity of Pfu DNA polymerase was investigated using the lacI-based PCR assay. The lacI mutational target gene was amplified from varying amounts of plasmid DNA and from a single-copy transgene in rodent genomic DNA, and error rates (mutation frequency per base pair per duplication) were found to be independent of the number of target duplications (data not shown). No significant difference in error rate was observed between PfuTurbo and Pfu DNA polymerases under several different PCR amplification conditions (table 1). In addition, the PCR enhancing factor did not alter the fidelity of Pfu DNA polymerase when added to PCR amplifications at concentrations ranging from 0.2- to 100-fold of the level present in PfuTurbo DNA polymerase (data not shown).

Table 1
Performance Comparisons of Stratagenes PCR Enzymes

Error rate*
(x10-6)

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