Development of a high-speed real-time PCR system for rapid and precise nucleotide recognition

Hideyuki Terazono, Hiroyuki Takei, Akihiro Hattori, Kenji Yasuda

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

Polymerase chain reaction (PCR) is a common method used to create copies of a specific target region of a DNA sequence and to produce large quantities of DNA. A few DNA molecules, which act as templates, are rapidly amplified by PCR into many billions of copies. PCR is a key technology in genome-based biological analysis, revolutionizing many life science fields such as medical diagnostics, food safety monitoring, and countermeasures against bioterrorism. Thus, many applications have been developed with the thermal cycling. For these PCR applications, one of the most important key factors is reduction in the data acquisition time. To reduce the acquisition time, it is necessary to decrease the temperature transition time between the high and low ends as much as possible. We have developed a novel rapid real-time PCR system based on rapid exchange of media maintained at different temperatures. This system consists of two thermal reservoirs and a reaction chamber for PCR observation. The temperature transition was achieved within 0.3 sec, and good thermal stability was achieved during thermal cycling with rapid exchange of circulating media. This system allows rigorous optimization of the temperatures required for each stage of the PCR processes. Resulting amplicons were confirmed by electrophoresis. Using the system, rapid DNA amplification was accomplished within 3.5 min, including initial heating and complete 50 PCR cycles. It clearly shows that the device could allow us faster temperature switching than the conventional conduction-based heating systems based on Peltier heating/cooling.

Original languageEnglish
Title of host publicationAdvanced Environmental, Chemical, and Biological Sensing Technologies VII
Volume7673
DOIs
Publication statusPublished - 2010
Externally publishedYes
EventAdvanced Environmental, Chemical, and Biological Sensing Technologies VII - Orlando, FL, United States
Duration: 2010 Apr 52010 Apr 6

Other

OtherAdvanced Environmental, Chemical, and Biological Sensing Technologies VII
CountryUnited States
CityOrlando, FL
Period10/4/510/4/6

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Keywords

  • bioterrorism
  • circulating water
  • DNA sequencing
  • Droplet
  • heat-exchange
  • high speed
  • medical diagnostics
  • PCR
  • Polymerase Chain Reaction

ASJC Scopus subject areas

  • Applied Mathematics
  • Computer Science Applications
  • Electrical and Electronic Engineering
  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics

Cite this

Terazono, H., Takei, H., Hattori, A., & Yasuda, K. (2010). Development of a high-speed real-time PCR system for rapid and precise nucleotide recognition. In Advanced Environmental, Chemical, and Biological Sensing Technologies VII (Vol. 7673). [76730U] https://doi.org/10.1117/12.849819