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Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR,,,Performance

s followed by 10 cycles at 95C for 30 seconds (denaturation), 58C for 30 seconds (annealing), and 72C for 2 minutes; followed by 20 cycles at 95C for 30 seconds, 58C for 30 seconds, and 72C for 2 minutes plus an additional 5 seconds per cycle. The conditions for RT-PCR (Figure 4) were: (RoboCycler 96 temperature cycler) 1 cycle at 95C for 1 minute followed by 30 cycles at 95C for 1 minute (denaturation), 60C for 1 minute (annealing), and 68C for 1 minute (targets <1 kb) or 2 minutes per kb (targets >4 kb). For each cycling regimen, one final extension cycle of 72C for 7 minutes was performed (after 30 cycles).

PCR products were electrophoresed on 1 to 4% agarose/1X TBE gels. PCR products were visualized by ethidium bromide staining, and the gels were imaged using the Eagle Eye II still video system.

REFERENCES

  1. Borns, M. and Hogrefe, H. (2000) Strategies 13: 1-3.

  2. Barnes, W.M. (1994) Proc. Natl. Acad. Sci. 91: 2216-2220.

  3. Borns, M., et al. (1999) Strategies 12: 33-36.

  4. Landre, P.A., Gelfand, D.H., and Watson, R.M. (1995) In PCR Strategies. Academic Press, San Diego, pp. 3-16.

* U.S. Patent Nos. 5,545,552, 5,556,772, 5,866,395 and 5,948,663 and patents pending.


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