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Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase

ymerase was shown to successfully amplify vector targets up to 10 kb using 1-minute-per-kb extension times (lanes 13, 15 and 17) and a 14-kb lambda DNA target using 2-minute-per-kb extension times (lane 19). When relatively short vector targets were amplified, the most dramatic differences in PCR product yield between PfuTurbo and Pfu DNA polymerases were observed when limiting amounts of DNA template were used. For example, PfuTurbo DNA polymerase produced significantly more 1.9-kb PCR product when amplification was performed for 30 cycles using 1 pg of plasmid DNA (lane 13). PfuTurbo and Pfu DNA polymerases generated comparable yields of product when 10 pg of plasmid DNA was used (data not shown). Dramatic differences in yield between PfuTurbo and Pfu DNA polymerases were also observed when amplifying longer vector targets. For example, Pfu DNA polymerase amplified an 8-kb target from lambda phage DNA (lane 16), but not a 14-kb lambda DNA target (lane 20) nor a 10-kb, 40-copy amplicon from transgenic mouse genomic DNA (lane 18). In contrast, PfuTurbo DNA polymerase synthesized high yields of all of the longer vector targets (lanes 15, 17 and 19).

In total, the data demonstrate that PCR product yields generated with PfuTurbo DNA polymerase are higher than those generated with Pfu DNA polymerase (figure 2) or with other single PCR enzymes (Figure 1). PfuTurbo DNA polymerase can be used routinely in all PCR applications that utilize Taq DNA polymerase or a proof- reading PCR enzyme (e.g., Pfu, VentR, Deep VentR, Pwo, KOD and Ultma DNA polymerases). PfuTurbo DNA polymerase is particularly suited for amplifying longer (>2 kb) or complex genomic DNA targets, which are not readily amplified by single D
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