High speed polymerase chain reaction in constant flow

Biosci Biotechnol Biochem. 1994 Feb;58(2):349-52. doi: 10.1271/bbb.58.349.

Abstract

A new simple reactor of the tubing type was developed for polymerase chain reaction (PCR). A thin Teflon capillary tube was used as a tubing reactor in which the reaction mixture of PCR was driven by a pump at a constant flow rate. The sample was treated with three successive thermal stages for denaturation, annealing, and elongation of DNA and primers as a function of the position in the tube. The amplification yield was about a half of that obtained by a commercial thermocycler. Moreover, the total reaction time from 12 to 18 min, which was one-tenth of the time generally required by conventional thermocyclers using metal blocks, assured substaintial amplification of a DNA fragment. In addition, this reactor could be also used for rapid cycle-sequences. This new device will be easily incorporated into automated and rapid DNA analysis systems for DNA sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Bacterial / analysis
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques
  • Polymerase Chain Reaction / instrumentation*
  • Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA / instrumentation*
  • Sequence Analysis, DNA / methods
  • Templates, Genetic
  • Time Factors

Substances

  • DNA, Bacterial