Development of a high-speed real-time polymerase chain reaction system using a circulating water-based rapid heat-exchange

Hideyuki Terazono, Hiroyuki Takei, Akihiro Hattori, Kenji Yasuda

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9 Citations (Scopus)

Abstract

Polymerase chain reaction (PCR) is a powerful technique to detect microorganisms, viruses, or cells by amplifying a single copy or a few copies of a fragment of a particular DNA sequence. To reduce acquisition time, it is necessary to decrease the temperature transition time between denaturation and extension. We have developed a simple rapid real-time microlitter-sample droplet PCR system accomplished by the rapid liquidbased heat-exchange of sample droplets by quick switching of two circulating hot waters of denaturation and extension, a microlitter-sized droplet and a thin-film aluminum chip. Using this system, rapid PCR amplification of a set of droplets lined up on an aluminum chip was conducted successfully as shown by the increase in fluorescence intensity, and was accomplished within 3.5 min in 40 cycles of 1 s denaturation and 3 s extension reaction, which is one magnitude faster than conventional fast PCR systems. This method allows the rapid detection of DNA fragments and has a possibility for measuring multiple samples simultaneously in a miniaturized microfluidic chip.

Original languageEnglish
JournalJapanese Journal of Applied Physics
Volume49
Issue number6 PART 2
DOIs
Publication statusPublished - 2010 Jun
Externally publishedYes

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ASJC Scopus subject areas

  • Engineering(all)
  • Physics and Astronomy(all)

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