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Easily Amplify Genomic DNA with Long-Distance PCR

directly for PCR amplification.

Conclusions

This procedure allows fast genotyping of a large number of mice with long-distance PCR using TaqPlus Long DNA polymerase. Usually, the results are ready the day after mice ear clipping. Additionally, by amplifying several specific overlapping fragments and analyzing these fragments, a more detailed characterization of the region of interest can be realized. This procedure will have wide application in transgenic mice technology and may be useful for detailed analysis of chromosomal loci with extensive modifications, either introduced by homologous recombination or naturally occurring (i.e., deletions and insertions).

REFERENCES

  1. Abbott, C., et al. (1988) Trends Genet. 4: 325.

  2. Hanley, T. and Merlie, J.P. (1991) Biotechniques 10: 56.

  3. Winberg, G. (1991) PCR Methods Appl. 1: 72-74.

  4. Hogan, B., Constantini, F., and Lacy, E. (1986) Manipulating the Mouse Embryo. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

  5. Hasty, P. and Bradley, A. (1995) In: Joyner, A.L. ed., Gene Targeting, A Practical Approach. Oxford University Press, New York. pp. 1-31.

  6. Kim, H.-S. and Smithies, O. (1988) Nucleic Acids Res. 16: 8887-8903.

  7. Cheng, S., et al. (1995) PCR Methods Appl. 4: 294-298.

  8. Nadon, N.L. and Draeger, K. (1996) Transgenic Res. 5: 209-211.

* U.S. Patent No. 5,556,772 and patents pending.


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