( Fig. 3: TripleMaster PCR System in ...)The TripleMaster PCR System for three different amplifications

Fig. 3: TripleMaster PCR System in a test with a GC-rich template Conclusions

The exceptionally high extension rate of the innovative enzyme mix (consisting of high-quality Taq polymerase, polymerase enhancing factor, and proofreading enzyme) combines with the new buffer systems (for long and short range) to produce an impressive increase in yields for long PCR.

As the accuracy of this new system reaches a level normally reserved for highly specific kits that can only be used for highfidelity PCR, the TripleMaster PCR System is an all-purpose tool suitable not only for high-fidelity experiments, but for other applications as well.

For GC-rich templates (the secondary structures which often cause complex applicational problems) adding low concentrations of DMSO or Eppendorf TaqMaster is sufficient to obtain very good results. Since TaqMaster, unlike DMSO, has no negative effect on the fidelity of Taq DNA polymerase (data not shown) it is an ideal alternative particularly for subsequent cloning and sequencing reactions.

Literature

[1] Barnes, W. M.; PCR amplification of up to 35 kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA. 1994 Mar 15; 91 (6): 2216-2220.

[2] Cheng, S., Kolmodin, L.A.; XL PCR amplification of long targets from genomic DNA. Methods-Mol-Biol. 1997; 67: 17-29.

[3] Wagner, R. and Dean, A. 1998. The use of immobilizes mismatch binding protein for optimization of PCR fidelity. In: PCR Methods Manual (Innis, M., Gelfand, D. and Sninsky, J., eds).



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