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Comparing Fidelity and Performance of Proofreading PCR Enzymes


PfuTurbo DNA polymerase is superior to Platinum Pfx DNA polymerase

Michael Borns Janice Cline Holly Hogrefe
Stratagene

We recently evaluated the properties of Platinum Pfx DNA polymerase, a competitors new PCR enzyme formulation consisting of KOD DNA polymerase and polymerase-specific antibodies. Comparisons revealed that Stratagenes PfuTurbo DNA polymerase *, is superior to Pfx DNA polymerase, with respect to both replication fidelity and PCR performance. The intrinsic error rate of Pfx DNA polymerase was found to be 2.7-fold higher than the average error rate of PfuTurbo DNA polymerase. Moreover, in side-by-side comparisons, PfuTurbo DNA polymerase provided higher product yields, greater sensitivity, and amplification of longer targets than Pfx DNA polymerase.

The development of high-fidelity PCR enzymes for accurate DNA replication has greatly simplified a number of laboratory procedures, including cloning, directed mutagenesis, and mutation detection. The enzyme best suited for high-fidelity PCR applications is Stratagenes Pfu DNA polymerase, the most accurate thermostable DNA polymerase described to date.1-3 PCR reaction conditions have been optimized for highest fidelity,1 and further improvements in PCR product yield, sensitivity, rate, and target-length capability have been achieved with the discovery of a novel PCR-enhancing factor**, referred to as the Turbo enhancer.4 PCR comparisons have demonstrated that PfuTurbo DNA polymerase amplifies longer targets in higher yield than Taq DNA polymerase, cloned Pfu DNA polymerase (without enhancer), and other Pfu-related proofreading enzymes, including Vent, Deep Vent, KOD, and Pwo DNA polymerases.4,5

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TAG: Comparing Fidelity and Performance Proofreading PCR Enzymes

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dNTP Set from Invitrogen
StrepMAB-Immo coated microplate; 96 well from IBA GmbH
cAMP[125I] Direct SPA Screening Biotrak Assay from GE Healthcare, formerly Amersham Biosciences
GIBCO Leibovitz's L-15 Medium powder from Invitrogen
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