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Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase

DNA polymerase has been shown to be ideally suited for both high-fidelity and higher-throughput PCR applications. In this article, we demonstrate that PCR product yields and replication fidelity are significantly higher with PfuTurbo DNA polymerase than with other PCR enzymes.

PCR Performance of PfuTurbo DNA Polymerase

Figure 1 shows the results of amplifying 0.9-kb and 5.2-kb targets from human genomic DNA using PfuTurbo DNA polymerase and several other PCR enzymes. For these PCR amplifications, extension times of 1 minute per kb of target and varying amounts of each polymerase (from 0.5 to 5 U) were used. All of the enzymes tested successfully amplified the 0.9-kb target. The reaction products shown in lanes 1-7 were generated using 2 to 2.5 U of each DNA polymerase, with the exception that KOD DNA polymerase required 1.25 U to obtain sufficient yields with minimal smearing. Higher yields of PCR products were obtained when 5 U of Pfu, Pwo or Taq DNA polymerase was used (data not shown). Smearing or reduced yield was observed when higher amounts of KOD, VentR or Deep VentR DNA polymerases were used (data not shown).

The 5.2-kb target was also amplified with varying amounts (from 0.5 to 5 U) of each DNA polymerase. The reactions shown in Figure 1 (lanes 8-14) are those that exhibited the highest yields of PCR products (for PfuTurbo and Taq DNA polymerases) or those that were carried out using the highest possible number of polymerase units before excessive smearing was observed. Only PfuTurbo (lane 8) and Taq (lane 14) DNA polymerases successfully amplified the 5.2-kb target, and
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