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High-Fidelity PCR with a Novel Polymerase Mixture

rate of Pfu DNA polymerase, but 1.9-fold and 2.1-fold lower than the error rates of the TaqPlus Long PCR system and Taq DNA polymerase, respectively. The TaqPlus Precision reaction buffer, provided in this system is also critical for optimal PCR fidelity (data not shown). Moreover, the TaqPlus Precision PCR system was found to exhibit the highest average replication accuracy that has been achieved using commercially available DNA polymerase mixtures.25

Figure 2

The TaqPlus Precision PCR system also tends to generate higher yields of PCR product than can be obtained with either Taq DNA polymerase or Pfu DNA polymerase. Figure 2 shows a typical comparison of PCR product yields for Taq DNA polymerase and the TaqPlus Precision PCR system for various target systems. The TaqPlus Precision PCR system successfully synthesizes vector PCR targets up to 15 kb in length and genomic PCR targets up to 10 kb in length using a relatively short extension time of 1 minute per kb of DNA target.

Comparison of PCR Performance

Table 1 summarizes the recommended PCR extension time, error rate, relative PCR product yield and useful target length for Stratagene's four thermophilic DNA polymerases and polymerase mixtures. Taq DNA polymerase is relatively fast and reliable and is suitable for amplifying targets 4 kb and smaller when fidelity is not a concern. Pfu DNA polymerase has unusually high thermostability and the lowest error rate of any thermostable DNA polymerase studied to date. Clearly, Pfu DNA polymerase is the enzyme of choice for PCR a
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