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PubMed · 15706574

High-throughput single-strand conformation polymorphism analysis on a microfabricated capillary array electrophoresis device.

Huijun Tian·Charles A Emrich·James R Scherer·Richard A Mathies·Paal Skytt Andersen·Lars Allan Larsen·Michael Christiansen
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Abstract

A high-density 384-lane microfabricated capillary array electrophoresis device is evaluated for high-throughput single-strand conformation polymorphism (SSCP) analysis. A delayed back bias direct electrokinetic injection scheme is used to provide better than 10-bp resolution with an 8.0-cm effective separation length. Separation of a HaeIII digest of PhiX174 yielded theoretical plate numbers of 4.0 x 10(6). Using 5% PDMA containing 10% glycerol and 15% urea, 21 single-nucleotide polymorphisms (SNPs) from HFE, MYL2, MYL3, and MYH7 genes associated with hereditary hemochromatosis (HHC) and hereditary hypertrophic cardiomyopathy (HCM) are discriminated at two running temperatures (25 degrees C and 40 degrees C), providing 100% sensitivity. The data in this study demonstrate that the 384-lane microCAE device provides the resolution and detection sensitivity required for SSCP analysis, showing its potential for ultrahigh-throughput mutation detection.

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CardiomyopathiesDNA Mutational AnalysisElectrophoresis, CapillaryHemochromatosisHumansMicrochemistryPolymorphism, Single-Stranded ConformationalEvaluation StudyJournal ArticleResearch Support, N.I.H., ExtramuralResearch 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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Huijun Tian, Charles A Emrich, James R Scherer, Richard A Mathies, Paal Skytt Andersen, Lars Allan Larsen, Michael Christiansen. 2005. High-throughput single-strand conformation polymorphism analysis on a microfabricated capillary array electrophoresis device.. https://doi.org/10.1002/elps.200410205

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