Document Detail


Alternative method of allelic discrimination.
MedLine Citation:
PMID:  25109294     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
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.
Authors:
Rajeswari Avula; Dennis O'Kane; John L Black
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Publication Detail:
Type:  Journal Article     Date:  2014-08-01
Journal Detail:
Title:  BioTechniques     Volume:  57     ISSN:  1940-9818     ISO Abbreviation:  BioTechniques     Publication Date:  2014  
Date Detail:
Created Date:  2014-08-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  England    
Other Details:
Languages:  eng     Pagination:  88-90     Citation Subset:  IM    
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