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Related Experiment Videos

High speed polymerase chain reaction in constant flow

H Nakano1, K Matsuda, M Yohda

  • 1Department of Applied Biological Sciences, School of Agriculture, Nagoya University, Japan.

Bioscience, Biotechnology, and Biochemistry
|February 1, 1994
PubMed
Summary

A novel tubing reactor significantly speeds up polymerase chain reaction (PCR) DNA amplification, reducing reaction times to 12-18 minutes. This innovation offers substantial amplification for DNA fragments and rapid cycle sequencing applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Conventional thermocyclers using metal blocks are time-consuming for polymerase chain reaction (PCR).
  • There is a need for faster and more efficient DNA amplification methods for various applications.
  • Integrating PCR into automated DNA analysis systems requires rapid and compact reactor designs.

Purpose of the Study:

  • To develop a simple, tubing-type reactor for rapid polymerase chain reaction (PCR).
  • To evaluate the performance of the tubing reactor in terms of amplification yield and reaction time.
  • To assess the potential of the reactor for automated DNA analysis and DNA sequencing.

Main Methods:

  • Utilized a thin Teflon capillary tube as the reactor.

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  • Employed a pump to drive the PCR reaction mixture at a constant flow rate.
  • Implemented three successive thermal stages (denaturation, annealing, elongation) along the tube's length.

    • Main Results:

    • Achieved substantial amplification of DNA fragments within 12 to 18 minutes.
    • Reaction time was approximately one-tenth of that required by conventional thermocyclers.
    • Amplification yield was approximately 50% of that obtained with a commercial thermocycler.

    Conclusions:

    • The developed tubing reactor offers a significant reduction in PCR reaction time.
    • The device is suitable for rapid DNA amplification and rapid cycle sequencing.
    • This reactor can be readily integrated into automated DNA analysis systems for applications like DNA sequencing.