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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 kbPlus 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 kbPlus 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 kbPlus 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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All-in-one mix, removes primers and dNTPs from PCR product for following direct sequencing...
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