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

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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Figure 1: Hot-stop PCR.
Figure 2: Comparison of regular PCR and hot-stop PCR for quantitation of allele ratios.

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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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Correspondence to Andrew 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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