Microchip Electrophoresis for Specific Gene Detection of the Pathogenic Bacteria V. cholerae by Circle-to-Circle Amplification

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  • MAHMOUDIAN Laili
    Department of Applied Chemistry, Graduates School of Engineering, Nagoya University
  • MELIN Jonas
    Department of Genetic and Pathology, Uppsala University, Rudbeck Laboratory Biomedical Diagnostic Institute, Dublin City University
  • MOHAMADI Mohamad Reza
    Laboratory of Macromolecules and Microsystems in Biology and Medicine (MMBM), Department of Physical Chemistry, Institute Curie
  • YAMADA Keiko
    Department of Molecular Biology, Graduate School of Medicine, Nagoya University
  • OHTA Michio
    Department of Molecular Biology, Graduate School of Medicine, Nagoya University
  • KAJI Noritada
    Department of Applied Chemistry, Graduates School of Engineering, Nagoya University MEXT Innovative Research Center for Preventive Medical Engineering, Nagoya University
  • TOKESHI Manabu
    Department of Applied Chemistry, Graduates School of Engineering, Nagoya University MEXT Innovative Research Center for Preventive Medical Engineering, Nagoya University
  • NILSSON Mats
    Department of Genetic and Pathology, Uppsala University, Rudbeck Laboratory
  • BABA Yoshinobu
    Department of Applied Chemistry, Graduates School of Engineering, Nagoya University MEXT Innovative Research Center for Preventive Medical Engineering, Nagoya University Plasma Nanotechnology Research Center, Nagoya University Health Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST) Institute for Molecular Science, National Institutes of Natural Sciences

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We have developed a new method for a fast and precise analysis of circle-to-circle amplification (C2CA) product for specific gene detection by microchip electrophoresis. In this method, we have added a new enzymatic step to the C2CA reaction, which could be carried out isothermally at 37°C. Compared to the original single-stranded DNA, the double-stranded DNA that is produced by this enzymatic reaction is more reliable for analysis by microchip electrophoresis. C2CA product was detected within 55 s with high reproducibility by this method which was successfully applied to the detection of 10-ng genomic DNA of the pathogenic bacteria Vibrio. cholerae within 110 s. Purification was found to be an indispensable step for the analysis of the C2CA product of genomic DNA samples.

収録刊行物

  • Analytical Sciences

    Analytical Sciences 24 (3), 327-332, 2008

    社団法人 日本分析化学会

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