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Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase

PCR product yields were significantly higher with PfuTurbo DNA polymerase. None of the other DNA polymerases could amplify the 5.2-kb genomic target despite the use of up to 5 U of enzyme or longer extension times (data not shown).

Lack of amplification of the 5.2-kb target by Pfu, Pwo, VentR, Deep VentR and KOD DNA polymerases was unrelated to the particular primer-template system used. None of these enzymes, including Taq DNA polymerase, could amplify a 6.6-kb target using a different primer set and mouse genomic DNA as template (data not shown). PfuTurbo DNA polymerase, however, was able to amplify this target.

figure 2

PfuTurbo DNA polymerase has been used to amplify a variety of DNA targets, ranging in complexity from plasmid and lambda phage DNA templates to complex, single-copy amplicons from genomic DNA. The data in figure 2 demonstrate that PfuTurbo DNA polymerase synthesizes higher yields of PCR product than Pfu DNA polymerase alone, regardless of the primer-template system used. PfuTurbo DNA polymerase successfully amplified complex genomic targets of 0.9 to 5.2 kb in length using 1-minute-per-kb extension times (lanes 1-6) and genomic targets of 6.1 to 10 kb in length using 2-minute-per-kb extension times (lanes 7-12). Under the same conditions, Pfu DNA polymerase successfully amplified only the 0.9- and 1.9-kb targets (lanes 2 and 4), and product yields were significantly lower than those obtained using PfuTurbo DNA polymerase (lanes 1 and 3). To achieve higher yields of the 0.9- and 1.9-kb targets with Pfu DNA polymerase, greater amounts of polymerase or longer extension times (2 minutes per kb of target) must be used (data not shown).

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