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Related Experiment Videos

Microchip device for cell lysis, multiplex PCR amplification, and electrophoretic sizing

L C Waters1, S C Jacobson, N Kroutchinina

  • 1Oak Ridge National Laboratory, Tennessee.

Analytical Chemistry
|February 14, 1998
PubMed
Summary
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This study presents a microchip that integrates cell lysis, multiplex polymerase chain reaction (PCR) amplification, and rapid electrophoretic analysis for DNA. This streamlined process enables quick DNA analysis from biological samples.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Traditional DNA analysis involves multiple complex steps.
  • Microfluidic devices offer potential for integrated biological sample processing.
  • Rapid and efficient DNA amplification and separation are crucial for diagnostics.

Purpose of the Study:

  • To develop and demonstrate a monolithic microchip for integrated cell lysis, multiplex PCR, and electrophoretic DNA analysis.
  • To achieve rapid DNA amplification and size separation on a single device.
  • To validate the microchip's performance using specific DNA targets.

Main Methods:

  • Sequential execution of cell lysis, multiplex PCR amplification, and electrophoretic analysis on a monolithic microchip.
  • Thermal cycling for cell lysis and DNA amplification.

Related Experiment Videos

  • Size separation using sieving media (hydroxyethyl cellulose or poly(dimethylacrylamide)) and fluorescence detection with an intercalating dye.
  • Analysis of amplified DNA fragments from bacteriophage lambda and E. coli.

    • Main Results:

    • Successful integration of cell lysis, multiplex PCR, and electrophoresis on a single microchip.
    • Amplification of specific DNA regions (500 bp from bacteriophage lambda; 154, 264, 346, 410, 550 bp from E. coli).
    • Electrophoretic analysis completed in under 3 minutes post-amplification.
    • Accurate DNA sizing demonstrated using a DNA sizing ladder.

    Conclusions:

    • The developed microchip enables rapid, integrated DNA analysis.
    • This technology has potential applications in molecular diagnostics and high-throughput screening.
    • The microchip platform offers a streamlined approach to complex molecular biology workflows.