Denaturing gradient-based two-dimensional gene mutation scanning in a polymer microfluidic network

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

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

An integrated two-dimensional (2-D) DNA separation platform, combining standard gel electrophoresis with temperature gradient gel electrophoresis (TGGE) on a polymer microfluidic chip, is reported. Rather than sequentially sampling DNA fragments eluted from standard gel electrophoresis, size-resolved fragments are simultaneously electrokinetically transferred into an array of orthogonal microchannels and screened for the presence of sequence heterogeneity by TGGE in a parallel and high throughput format. A bulk heater assembly is designed and employed to externally generate a temporal temperature gradient along an array of TGGE channels. Extensive finite element modeling is performed to determine the optimal geometries of the microfluidic network for minimizing analyte band dispersion caused by interconnected channels in the network. A pH-mediated on-chip analyte stacking strategy is employed prior to the parallel TGGE separations to further reduce additional band broadening acquired during the electrokinetic transfer of DNA fragments between the first and second separation dimensions. A comprehensive 2-D DNA separation is completed in less than 5 min for positive detection of single-nucleotide polymorphisms in multiplex PCR products that vary in size and sequence.

Original languageEnglish (US)
Pages (from-to)392-400
Number of pages9
JournalLab on a Chip
Volume5
Issue number4
DOIs
StatePublished - Apr 2005

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Chemistry(all)
  • Biomedical Engineering

Fingerprint Dive into the research topics of 'Denaturing gradient-based two-dimensional gene mutation scanning in a polymer microfluidic network'. Together they form a unique fingerprint.

  • Cite this

    Buch, J. S., Rosenberger, F., Highsmith, W. E., Kimball, C., DeVoe, D. L., & Lee, C. S. (2005). Denaturing gradient-based two-dimensional gene mutation scanning in a polymer microfluidic network. Lab on a Chip, 5(4), 392-400. https://doi.org/10.1039/b416682e