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Efficiently Insert Unique Restriction Sites into Plasmid Vectors

d Xho I restriction digestion patterns for seven of the clones are shown in Figure 2, indicating that the appropriate restriction sites were successfully introduced into the plasmid vector.

Fig.2

Conclusions

These results demonstrate that a 28-base insertion, encoding three unique restriction enzyme sites, was effectively introduced into a plasmid vector using the QuikChange mutagenesis kit. Mutagenesis reactions and transformation can be completed in less than one day so putative positive clones may be examined the following day. And, because high-fidelity PfuTurbo DNA polymerase is used, random mutations are minimized, rendering this the preferred method for efficient mutagenesis.

REFERENCES

  1. Papworth, C., Bauer, J.C., and Braman, J. (1996) Strategies 9: 3-4.

  2. Hogrefe, H., et al. (1997) Strategies 10: 93-96.

  3. Braman, J. and Basehore, S. (1997) Strategies 10: 84-86.

* U.S. Patent No. 5,789,166 and patents pending
** U.S. Patent Nos. 5,545,552 and 5,866,395 and patents pending


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