( m Taq DNA polymerase High Fidelity (L...)Unique DNA Polymerase Formulation Excels in a Broad Range of PCR,,,Applications

m Taq DNA polymerase High Fidelity (Life Technologies, Inc.), a blend developed specifically for high-fidelity PCR applications, and the Expand 20 kb^Plus PCR system (Roche Molecular Biochemicals), the only commercial blend specifically formulated for amplifications of extra-long (>20 kb) genomic targets. The range of amplicon lengths synthesized by each formulation was evaluated by amplifying a panel of short genomic DNA targets, ranging in length from 0.1 kb to 0.5 kb (Figure 1) and a panel of extra-long targets, ranging in length from 17 kb to 48 kb (Figure 2). Amplifications were performed under identical conditions (Methods) with the following exceptions: as per the manufacturers recommendations, PCRs were carried out using 2.5 U of Platinum Taq DNA polymerase High Fidelity, 2.6 U of Expand 20 kb^Plus PCR system blend, and 2.5 U (<1 kb) or 5 U (>17 kb) of Herculase enhanced DNA polymerase. The 30-kb and 48-kb targets were amplified with the Herculase formulation in the presence of 3% and 7% DMSO, respectively.

Fig.2

The Platinum Taq DNA polymerase High Fidelity exhibited limited target-length capability compared to Herculase enhanced DNA polymerase, which successfully amplified all targets up to 48 kb in length (Figure 1 and Figure 2). Even in amplifications of short targets, the Herculase formulation produced higher product yields than the Platinum Taq blend (Figure 1). Although the Expand 20 kb^Plus PCR system successfully amplified the longer 17-kb and 30-kb genomic targets, product yields were highest with the Herculase formulation. Moreover, amplification of the longest 4
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