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

required for routine PCRs, we recommend adding DMSO (3 to 7% final concentration) to amplifications of genomic targets greater than 23 kb and lambda targets greater than 30 kb to increase product yield.1 DMSO can also be used to facilitate amplification of GC-rich targets and amplicons containing difficult secondary structures that impede DNA polymerase translocation.4 Such targets can be difficult to amplify in high yield.

Fig.5

We tested the utility of Herculase DNA polymerase in amplifying GC-rich targets (Figure 5). A 300-bp amplicon with 82.5% GC content was amplified with or without DMSO using Herculase or Taq DNA polymerase. Successful amplification of this extremely difficult target required modification of the initial denaturation step, whereby the initial denaturation temperature was increased from 95C to 98C, and the incubation time was increased from 1 minute to 3 minutes. Under these modified conditions, Herculase DNA polymerase successfully amplified the target in the presence of 6 to 9% DMSO, with optimal yields achieved using 8 to 9% DMSO. In contrast, Taq DNA polymerase was unable to amplify this target despite using optimal denaturation conditions and DMSO concentrations up to 9% (final concentration) (Figure 5).

Additional studies show that Herculase DNA polymerase plus DMSO permits superior performance compared to commercial PCR systems developed specifically for amplifying GC-rich targets. In Figure 5, higher product yields were achieved using Herculase DNA polymerase plus DMSO compared to the GC-RICH PCR System (Roche Molecular Biochemicals). For the GC-RICH PCR System, amplifications were performed as recommended, using a Taq-based DN
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