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High-Fidelity PCR with a Novel Polymerase Mixture

mended for relatively short target (010 kb) amplifications, while the high-salt buffer is optimal for longer target (535 kb) amplifications. The TaqPlus Long PCR system typically generates higher yields of both short and long PCR products than can be obtained using either component enzyme alone.21 Including Taq2000 DNA polymerase in the TaqPlus Long polymerase formulation ensures that artifactual background smearing is minimal when long extension times (8 minutes) are used.

As with many commercially available two-polymerase mixtures, a potential limitation of the TaqPlus Long PCR system is its relatively low fidelity compared to Pfu DNA polymerase (figure 1). The TaqPlus Long PCR system exhibits an error rate similar to Taq DNA polymerase, the predominant component in the TaqPlus Long polymerase mixture. To address the needs of researchers requiring both high yields of PCR product and high replication fidelity, Stratagene has added the TaqPlus Precison PCR system to its expanding line of PCR products.

Taqplus Precision PCR System for Highest Fidelity

The TaqPlus Precision PCR system consists of a unique mixture of Taq2000 DNA polymerase and cloned Pfu DNA polymerase and a PCR buffer, all optimized for highest PCR fidelity. Fidelity measurements of the TaqPlus Precision PCR system were carried out using a PCRbased forward mutation assay21,22 that utilizes the wellcharacterized lacIOZ target gene.23,24 figure 1 shows errorrate comparisons between Taq and Pfu DNA polymerases, the TaqPlus Long PCR system and the TaqPlus Precision PCR system. The TaqPlus Precision PCR system exhibits a mean error rate of 3.9 x 106 mutations per bp per duplication, which is 3-fold higher than the error
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