Hot-stop PCR: a simple and general assay for linear quantitation of allele ratios.
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.
Uejima
H
H
Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Lee
M P
MP
Cui
H
H
Feinberg
A P
AP
D000483:Alleles; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D018392:Genomic Imprinting; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D010761:Phosphorus Radioisotopes; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D010761:Phosphorus Radioisotopes; D007335:Insulin-Like Growth Factor II; D004247:DNA