Abstract − Analytical Sciences, 29(3), 367 (2013).
Parallel Real-Time PCR on a Chip for Genetic Tug-of-War (gTOW) Method
Toyohiro NAITO,*1,*2 Ai YATSUHASHI,*1 Noritada KAJI,*1,*2 Taeko ANDO,*3 Kazuo SATO,*4 Hisao MORIYA,*5 Hiroaki KITANO,*6 Takao YASUI,*1,*2 Manabu TOKESHI,*2,*7 and Yoshinobu BABA*1,*2,*8
*1 Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8603, Japan
*2 FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8603, Japan
*3 College of Science and Engineering Department of Micro System Technology, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan
*4 Department of Mechanical Engineering, Faculty of Engineering, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho, Toyota 470-0392, Japan
*5 Research Core for Interdisciplinary Sciences (RCIS), Okayama University, 3-1-1 Tsushima-Naka, Kita, Okayama 700-8530, Japan
*6 The Systems Biology Institute, Falcon Building 5F, 5-6-9 Shirokanedai, Minato, Tokyo 108-0071, Japan
*7 Division of Biotechnology and Macromolecular Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita, Sapporo 060-8628, Japan
*8 Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2217-14 Hayashi-cho, Takamatsu 761-0395, Japan
*2 FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8603, Japan
*3 College of Science and Engineering Department of Micro System Technology, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan
*4 Department of Mechanical Engineering, Faculty of Engineering, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho, Toyota 470-0392, Japan
*5 Research Core for Interdisciplinary Sciences (RCIS), Okayama University, 3-1-1 Tsushima-Naka, Kita, Okayama 700-8530, Japan
*6 The Systems Biology Institute, Falcon Building 5F, 5-6-9 Shirokanedai, Minato, Tokyo 108-0071, Japan
*7 Division of Biotechnology and Macromolecular Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita, Sapporo 060-8628, Japan
*8 Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2217-14 Hayashi-cho, Takamatsu 761-0395, Japan
A microchip-based real-time polymerase chain reaction (PCR) device has been developed for the genetic tug-of-war (gTOW) method that provides quantitative data for research on biorobustness and systems biology. The device was constructed of a silicon glass chip, a temperature controlling Peltier element, and a microscope. A parallel real-time amplification process of target genes on the plasmids and the housekeeping genes in a model eukaryote Saccharomyces cerevisiae were detected simultaneously, and the copy number of the target genes were estimated. The device provides unique quantitative data that can be used to augment understanding of the system-level properties of living cells.
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