Temperature-shift speed dependence of nonspecific amplification of polymerase chain reaction examined by 1480nm photothermal transition speed controllable high-speed polymerase chain reaction system

Hideyuki Terazono, Akihiro Hattori, Hyonchol Kim, Hiroyuki Takei, Fumimasa Nomura, Tomoyuki Kaneko, Kenji Yasuda

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We have examined the contribution of temperature shift speed from denaturation to extension for the reduction of nonspecific amplification caused by the mismatched primer-target attachment. We have newly developed the photothermal quantitative polymerase chain reaction (qPCR) system, in which the direct absorption of a 1480nm infrared laser beam was controlled by a rotating gradient neutral density (ND) filter to acquire the precise control of the desired speed of temperature shift between 60 and 95°C up to 1 s. The results showed that a quick shift of the temperature during the qPCR procedure reduced nonspecific amplicons with a significant reduction of qPCR time when we have chosen proper primer sets, whereas the non-proper primer set amplified nonspecific amplicons in the fast qPCR. The results indicate that the potential of quick qPCR using proper primers can reduce nonspecific amplification and the required time for qPCR measurement, and the necessity of more precise check of the matching of the primer template adequate for the fast temperature shift and for quick qPCR analysis.

Original languageEnglish
Article number06GK02
JournalJapanese Journal of Applied Physics
Volume52
Issue number6 PART 2
DOIs
Publication statusPublished - 2013 Jun
Externally publishedYes

Fingerprint

polymerase chain reaction
Polymerase chain reaction
primers
Amplification
high speed
shift
Temperature
temperature
Denaturation
Infrared lasers
biopolymer denaturation
reaction time
infrared lasers
Laser beams
attachment
templates
laser beams
filters
gradients

ASJC Scopus subject areas

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

Cite this

Temperature-shift speed dependence of nonspecific amplification of polymerase chain reaction examined by 1480nm photothermal transition speed controllable high-speed polymerase chain reaction system. / Terazono, Hideyuki; Hattori, Akihiro; Kim, Hyonchol; Takei, Hiroyuki; Nomura, Fumimasa; Kaneko, Tomoyuki; Yasuda, Kenji.

In: Japanese Journal of Applied Physics, Vol. 52, No. 6 PART 2, 06GK02, 06.2013.

Research output: Contribution to journalArticle

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AU - Yasuda, Kenji

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