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Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR,,,Performance

A polymerase blend and varying concentrations (0 to 2 M) of GC-RICH resolution solution (betaine). Amplifications were performed with the GC-RICH PCR System using both the RoboCycler 96 temperature cycler (amplification unsuccessful, data not shown) and the single-block cycler recommended by the manufacturer (Figure 5).

Herculase DNA polymerase was also compared to the Advantage-GC 2 PCR kit (CLONTECH). Amplification was carried out as recommended by the kits manufacturer using a hot start Taq DNA polymerase blend, a PCR buffer containing 5% DMSO, and various concentrations of GC-Melt reagent (0 to 1.5 M). Despite repeated attempts, we were unsuccessful in amplifying the 300-bp target using the Advantage-GC 2 PCR kit (data not shown).

Therefore, Herculase DNA polymerase can be used with the provided DMSO to amplify difficult GC-rich targets, obviating the need for specialized DNA polymerase blends and PCR additives.

Multiplex PCR

Fig.6

Herculase enhanced DNA polymerase was also tested in multiplex PCR applications. In Figure 6, Herculase DNA polymerase provided uniform amplification of three genomic DNA targets (300 bp, 500 bp, 900 bp). Using the same reaction conditions, greater variation in amplicon yield was observed with Taq DNA polymerase (data not shown).

PCR Fidelity Comparisons

Herculase enhanced DNA polymerase exhibits high fidelity due to its novel pfu-based composition. In previous studies,1 Herculase DNA polymerase was found to exhibit an average error rate of 2.8 x 10-6 mutation frequency per base pair per duplication.1 Comparisons show that the fidelity of Herculase DNA polymerase is comparable to (Vent, Deep
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