High-speed DNA genotyping using microfabricated capillary array electrophoresis chips

Anal Chem. 1997 Jun 1;69(11):2181-6. doi: 10.1021/ac961237+.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Benzothiazoles
  • DNA / genetics*
  • DNA / isolation & purification
  • DNA Fragmentation / genetics
  • Electrophoresis, Capillary*
  • Fluorescent Dyes / chemistry
  • Genotype
  • HLA Antigens / genetics*
  • Hemochromatosis / genetics*
  • Hemochromatosis Protein
  • Histocompatibility Antigens Class I / genetics*
  • Histocompatibility Testing / methods
  • Humans
  • Membrane Proteins*
  • Molecular Sequence Data
  • Quinolines
  • Thiazoles / chemistry

Substances

  • Benzothiazoles
  • Fluorescent Dyes
  • HFE protein, human
  • HLA Antigens
  • Hemochromatosis Protein
  • Histocompatibility Antigens Class I
  • Membrane Proteins
  • Quinolines
  • Thiazoles
  • thiazole orange
  • DNA

Associated data

  • GENBANK/Z92910