DNA Mutation Detection in a Polymer Microfluidic Network Using Temperature Gradient Gel Electrophoresis

Jesse S. Buch, Christopher Kimball, Frederick Rosenberger, W. Edward Highsmith, Don L. DeVoe, Cheng S. Lee

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

A miniaturized system for DNA mutation analysis, utilizing temperature gradient gel electrophoresis (TGGE) in a polycarbonate (PC) microfluidic device, is reported. TGGE reveals the presence of sequence heterogeneity in a given heteroduplex sample by introducing a thermal denaturing gradient that results in differences between the average electrophoretic mobilities of DNA sequence variants. Bulk heater assemblies are designed and employed to externally generate temperature gradients in spatial and temporal formats along the separation channels. TGGE analyses of model mutant DNA fragments, each containing a single base substitution, are achieved using both single-and 10-channel parallel measurements in a microfluidic plafform. Additionally, a comprehensive polymer microfluidic device containing an integrated microheater and sensor array is developed and demonstrated for performing spatial TGGE for DNA mutation analysis. The device consists of two PC modular substrates mechanically bonded together. One substrate is embossed with microchannels, and the other contains a tapered microheater, lithographically patterned along with an array of temperature sensors. Compared with the external heating approaches, the integrated platform provides significant reduction in power requirement and thermal response time while establishing more accurate and highly effective control of the temperature gradient for achieving improved separation resolution.

Original languageEnglish (US)
Pages (from-to)874-881
Number of pages8
JournalAnalytical Chemistry
Volume76
Issue number4
DOIs
StatePublished - Feb 15 2004

ASJC Scopus subject areas

  • Analytical Chemistry

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    Buch, J. S., Kimball, C., Rosenberger, F., Highsmith, W. E., DeVoe, D. L., & Lee, C. S. (2004). DNA Mutation Detection in a Polymer Microfluidic Network Using Temperature Gradient Gel Electrophoresis. Analytical Chemistry, 76(4), 874-881. https://doi.org/10.1021/ac034913y