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PubMed · 9183181

High-speed DNA genotyping using microfabricated capillary array electrophoresis chips.

A T Woolley·G F Sensabaugh·R A Mathies
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Abstract

Capillary array electrophoresis (CAE) chips have been designed and fabricated with the capacity to rapidly (< 160 s) analyze 12 different samples in parallel. Detection of all lanes with 0.3 s temporal resolution was achieved using a laser-excited confocal-fluorescence scanner. The operation and capabilities of these CAE microdevices were first determined by performing electrophoretic separations of pBR322 MspI DNA samples. Genotyping of HLA-H, a candidate gene for the diagnosis of hereditary hemochromatosis, was then performed to demonstrate the rapid analysis of biologically relevant samples. Two-color multiplex fluorescence detection of HLA-H genotypes was accomplished by prelabeling the standard pBR322 MspI DNA ladder with a red emitting bis-intercalation dye (butyl TOTIN) and on-column labeling of the HLA-H DNA with thiazole orange. This work establishes the feasibility of using CAE chips for high speed, high-throughput genotyping.

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BenzothiazolesDNADNA FragmentationElectrophoresis, CapillaryFluorescent DyesGenotypeHLA AntigensHemochromatosisHemochromatosis ProteinHistocompatibility Antigens Class IHistocompatibility TestingHumansMembrane ProteinsMolecular Sequence DataQuinolinesThiazolesJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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A T Woolley, G F Sensabaugh, R A Mathies. 1997-06-01. High-speed DNA genotyping using microfabricated capillary array electrophoresis chips.. https://doi.org/10.1021/ac961237%2B

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