Temperature-Shift Speed Dependence of Nonspecific Amplification of Polymerase Chain Reaction Examined by 1480 nm Photothermal Transition Speed Controllable High-Speed Polymerase Chain Reaction System

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概要

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• 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 1480 nm 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.

• 2013-06-25

著者

• Hattori Akihiro

  Institute of Biomaterials and Bioengineering Dept. of Biomedical Information, Tokyo Medical and Dent

• Yasuda Kenji

  Kanagawa Academy Of Science And Technology

• Kim Hyonchol

  Kanagawa Academy of Science and Technology

• Terazono Hideyuki

  Kanagawa Academy of Science and Technology

• Hattori Akihiro

  Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Chiyoda, Tokyo 101-0062, Japan

• Nomura Fumimasa

  Kanagawa Academy of Science and Technology, Kawasaki 213-0012, Japan

• Takei Hiroyuki

  Faculty of Life Sciences, Toyo University, Itakura, Gunma 374-0193, Japan

• Kaneko Tomoyuki

  Kanagawa Academy of Science and Technology, Kawasaki 213-0012, Japan

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