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High-Fidelity PCR with a Novel Polymerase Mixture

pplications requiring high fidelity or amplification through GCrich regions, when relatively long extension times can be used (2 minutes per kb of DNA target). Although Pfu DNA polymerase can amplify up to 25 kb, the yield of PCR product is generally lower than can be obtained with polymerase mixtures. The TaqPlus Long PCR system is ideally suited for generating high yields of PCR product and amplifying long targets up to 35 kb when fidelity is not the primary consideration. Finally, the TaqPlus Precision PCR system exhibits an error rate significantly lower than Taq DNA polymerase, the TaqPlus Long PCR system and other commercially available sbased polymerase mixtures.25 The TaqPlus Precision system is recommended for relatively rapid, highfidelity amplification of PCR targets up to 15 kb when Pfu DNA polymerase requires too long an extension time or yields are insufficient.

Conclusions

Cycling time, fidelity, target length and product yield are all important considerations when selecting a DNA polymerase or twopolymerase mixtures for PCR. Stratagene offers four different thermostable DNA polymerases and polymerase mixtures that are uniquely suited for particular PCR applications. The TaqPlus Precision PCR system, the latest addition to Stratagene's line of PCR products, offers researchers improved PCR fidelity.

REFERENCES

  1. Mullis, K.B., and Faloona, F.A. (1987) Methods Enzymol. 155: 335350.

  2. Hogrefe, H. Unpublished.

  3. Nielson, K.B., et al. (1996) Strategies 9: 78.

  4. Lundberg, K.S., et al. (1991) Gene 108: 16.

  5. Flaman, J.M., et al. (1994) Nucleic Acids Res. 22: 32593260.

  6. Cline
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