Hot-stop PCR: a simple and general assay for linear quantitation of allele ratios (original) (raw)

Nature Genetics volume 25, pages 375–376 (2000)Cite this article

A Correction to this article was published on 01 May 2001

Abstract

We have developed a simple, quantitative assay for measurement of allele ratios that circumvents the problem of heteroduplex formation skewing the results of restriction endonuclease digestion of PCR products. This assay, ‘hot-stop PCR’, involves addition of a radiolabelled PCR primer at the final cycle. We applied the assay to analysis of loss of imprinting (LOI) of the insulin-like growth factor II gene (IGF2) in tumours.

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References

  1. Lee, M.P., Hu, R.-J., Johnson, L.A. & Feinberg, A.P. Nature Genet. 15, 181–185 (1997).
    Article Google Scholar
  2. Qian, N. et al. Hum. Mol. Genet. 6, 2021–2029 (1997).
    Article CAS Google Scholar
  3. Cui, H., Horon, I.L., Ohlsson, R., Hamilton, S.R. & Feinberg, A.P. Nature Med. 4, 1276–1280 (1998).
    Article CAS Google Scholar
  4. Anonymous. ABI Prism DNA Sequencing Chemistry Guide (Perkin-Elmer, Foster City, 1995).
  5. Mansfield, E.S. Hum. Mol. Genet. 2, 43–50 (1993).
    Article CAS Google Scholar
  6. Ohlsson, R. et al. Genomic Imprinting: Causes and Consequences (Cambridge University Press, Cambridge, 1995).
    Google Scholar
  7. Mashal, R.D., Koontz, J. & Sklar, J. Nature Genet. 9, 177–183 (1995).
    Article CAS Google Scholar
  8. Takeda, S., Ichii, S. & Nakamura, Y. Hum. Mutat. 2, 112–117 (1993).
    Article CAS Google Scholar
  9. Pushnova, E.A. & Zhu, Y.S. Anal. Biochem. 260, 24–29 (1998).
    Article CAS Google Scholar
  10. Yun, K., Soejima, H., Merrie, A.E., McCall, J.L. & Reeve, A.E. J. Pathol. 187, 518–522 (1999).
    Article CAS Google Scholar
  11. Greenwood, A.D. & Burke, D.T. Genome Res. 6, 336–348 (1996).
    Article CAS Google Scholar
  12. Rainier, S. et al. Nature 362, 747–749 (1993).
    Article CAS Google Scholar
  13. Ogawa, O. et al. Nature 362, 749–751 (1993).
    Article CAS Google Scholar

Download references

Acknowledgements

We thank P. Onyango and J. Ravenel for discussions. This work was supported by grants from the National Institutes of Health to A.P.F.

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Author notes

  1. Hiroshi Uejima and Maxwell P. Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
    Hiroshi Uejima, Maxwell P. Lee, Hengmi Cui & Andrew P. Feinberg
  2. Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
    Hiroshi Uejima, Maxwell P. Lee, Hengmi Cui & Andrew P. Feinberg
  3. Department of Molecular Biology & Genetics and Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
    Andrew P. Feinberg

Authors

  1. Hiroshi Uejima
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  2. Maxwell P. Lee
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  3. Hengmi Cui
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  4. Andrew P. Feinberg
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Corresponding author

Correspondence toAndrew P. Feinberg.

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Uejima, H., Lee, M., Cui, H. et al. Hot-stop PCR: a simple and general assay for linear quantitation of allele ratios.Nat Genet 25, 375–376 (2000). https://doi.org/10.1038/78040

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