High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates [5]

Igor Medintz, Wendy W. Wong, Lorenzo Berti, Lawrence Shiow, Jennifer Tom, James Scherer, George Sensabaugh, Richard A. Mathies

Research output: Contribution to journalArticle

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Abstract

An assay is described for high-throughput single nucleotide polymorphism (SNP) genotyping on a microfabricated capillary array electrophoresis (CAE) microchip. The assay targets the three common variants at the HFE locus associated with the genetic disease hereditary hemochromatosis (HHC). The assay employs allele-specific PCR (ASPCR) for the C282Y (845g->a), H63D (187c->g), and S65C (193a->t) variants using fluorescently-labeled energy-transfer (ET) allele-specific primers. Using a 96-channel radial CAE microplate, the labeled ASPCR products generated from 96 samples in a reference Caucasian population are simultaneously separated with single-base-pair resolution and genotyped in under 10 min. Detection is accomplished with a laser-excited rotary four-color fluorescence scanner. The allele-specific amplicons are differentiated on the basis of both their size and the color of the label emission. This study is the first demonstration of the combined use of ASPCR with ET primers and microfabricated radial CAE microplates to perform multiplex SNP analyses in a clinically relevant population.

Original languageEnglish (US)
Pages (from-to)413-421
Number of pages9
JournalGenome Research
Volume11
Issue number3
DOIs
StatePublished - 2001

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Hemochromatosis
Capillary Electrophoresis
Single Nucleotide Polymorphism
Alleles
Mutation
Energy Transfer
Polymerase Chain Reaction
Color
Inborn Genetic Diseases
Base Pairing
Population
Lasers
Fluorescence

ASJC Scopus subject areas

  • Genetics

Cite this

High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates [5]. / Medintz, Igor; Wong, Wendy W.; Berti, Lorenzo; Shiow, Lawrence; Tom, Jennifer; Scherer, James; Sensabaugh, George; Mathies, Richard A.

In: Genome Research, Vol. 11, No. 3, 2001, p. 413-421.

Research output: Contribution to journalArticle

Medintz, I, Wong, WW, Berti, L, Shiow, L, Tom, J, Scherer, J, Sensabaugh, G & Mathies, RA 2001, 'High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates [5]', Genome Research, vol. 11, no. 3, pp. 413-421. https://doi.org/10.1101/gr.164701
Medintz, Igor ; Wong, Wendy W. ; Berti, Lorenzo ; Shiow, Lawrence ; Tom, Jennifer ; Scherer, James ; Sensabaugh, George ; Mathies, Richard A. / High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates [5]. In: Genome Research. 2001 ; Vol. 11, No. 3. pp. 413-421.
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