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

Nat Genet. 2000 Aug;25(4):375-6. doi: 10.1038/78040.


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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles*
  • DNA / genetics
  • DNA Primers
  • Genomic Imprinting
  • Humans
  • Insulin-Like Growth Factor II / genetics
  • Phosphorus Radioisotopes
  • Polymerase Chain Reaction / methods*


  • DNA Primers
  • Phosphorus Radioisotopes
  • Insulin-Like Growth Factor II
  • DNA