
doi: 10.2144/000114199
pmid: 25109294
5' nuclease assays for allelic discrimination use both a wild-type and a mutant probe. Here we present a new method for genotyping by 5' nuclease assays that dispenses with the mutant probe, using the wild-type probe together with a probe for a reference gene known to be present in two copies to determine the copy number of the wild type allele relative to the reference gene. The copy number of the wild-type allele then determines the genotype: two copies indicates homozygous wild-type; one copy indicates heterozygous; and zero copies indicates homozygous mutant. We were able to use our method to correctly genotype three alleles of the thiopurine methyl transferase (TPMT) gene: TPMT *2 (c.G238C), *3B (c.G460A) and *3C (c.A719G). Our approach can be used as an alternate allelic discrimination strategy that is cost effective when multiple TaqMan assays are performed on a sample.
thiopurine methyl transferase allelic discrimination, Deoxyribonucleases, DNA Copy Number Variations, Genotype, Genotyping Techniques, QH301-705.5, TaqMan, copy number variation, Methyltransferases, Humans, Biology (General), Alleles
thiopurine methyl transferase allelic discrimination, Deoxyribonucleases, DNA Copy Number Variations, Genotype, Genotyping Techniques, QH301-705.5, TaqMan, copy number variation, Methyltransferases, Humans, Biology (General), Alleles
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