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High speed single nucleotide polymorphism typing of a hereditary haemochromatosis mutation with capillary array electrophoresis microplates
Author(s) -
Medintz Igor,
Wong Wendy W.,
Sensabaugh George,
Mathies Richard A.
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20000701)21:12<2352::aid-elps2352>3.0.co;2-g
Subject(s) - amplicon , capillary electrophoresis , typing , microbiology and biotechnology , polymerase chain reaction , single nucleotide polymorphism , allele , biology , snp , genetics , chemistry , gene , genotype
A single nucleotide polymorphism (SNP) typing assay is developed and evaluated on a microfabricated capillary array electrophoresis system. Using fluorescently labeled allele‐specific primers, the S65C (193A→T) substitution associated with hereditary haemochromatosis in the HFE gene is genotyped. The covalently labeled polymerase chain reaction (PCR) products are separated on a microfabricated radial capillary array electrophoresis microplate using nondenaturing gel media in under two minutes. Detection is accomplished with a laser‐excited rotary confocal scanner. The Rox‐labeled A‐allele specific amplicon (211 bp) is differentiated from the R110‐labeled T‐allele specific amplicon (201 bp) by both size and color. This study demonstrates the feasibility of using allele‐specific PCR with covalently labeled primers for high speed fluorescent SNP typing on microfabricated radial capillary array electrophoresis microplates.