High speed single nucleotide polymorphism typing of a hereditary haemochromatosis mutation with capillary array electrophoresis microplates

A single nucleotide polymorphism (SNP) typing assay is developed and evalua ted on a microfabricated capillary array electrophoresis system. Using fluo rescently 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 separat ed on a microfabricated radial capillary array electrophoresis microplate u sing nondenaturing gel media in under two minutes. Detection is accomplishe d with a laser-excited rotary confocal scanner. The Rox-labeled A-allele sp ecific amplicon (211 bp) is differentiated from the R110-labeled T-allele s pecific amplicon (201 bp) by both size and color. This study demonstrates t he feasibility of using allele-specific PCR with covalently labeled primers for high speed fluorescent SNP typing on microfabricated radial capillary array electrophoresis microplates.