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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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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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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DOI: https://doi.org/10.1038/78040
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