Alternative method of allelic discrimination

Autor:	Dennis J. O'Kane, John L. Black, Rajeswari Avula
Rok vydání:	2014
Předmět:	Genetics Deoxyribonucleases DNA Copy Number Variations Genotype Genotyping Techniques Mutant Wild type Methyltransferases Biology General Biochemistry Genetics and Molecular Biology TaqMan Humans Copy-number variation Allele Gene Genotyping Alleles Biotechnology
Zdroj:	BioTechniques. 57:88-90
ISSN:	1940-9818 0736-6205
DOI:	10.2144/000114199
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1ad44eb5adc5f90e53066e253716c9a8 https://doi.org/10.2144/000114199 Zobrazit plný text záznamu