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

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Abstract

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.

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Authors and Affiliations

  1. Department of Applied Chemistry, Graduates School of Engineering, Nagoya University, Furo, Chikusa, Nagoya, 464-8603, Japan

    Laili Mahmoudian, Noritada Kaji, Manabu Tokeshi & Yoshinobu Baba

  2. Department of Genetic and Pathology, Uppsala University, Rudbeck Laboratory, se-75185, Uppsala, Sweden

    Jonas Melin & Mats Nilsson

  3. Biomedical Diagnostic Institute, Dublin City University, Dublin, Ireland

    Jonas Melin

  4. Laboratory of Macromolecules and Microsystems in Biology and Medicine (MMBM), Department of Physical Chemistry, Institute Curie, UMR 168, Paris, France

    Mohamad Reza Mohamadi

  5. Department of Molecular Biology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa, Nagoya, 466-8550, Japan

    Keiko Yamada & Michio Ohta

  6. MEXTInnovative Research Center for Preventive Medical Engineering, Nagoya University, Furo, Chikusa, Nagoya, 464-8603, Japan

    Noritada Kaji, Manabu Tokeshi & Yoshinobu Baba

  7. Plasma Nanotechnology Research Center, Nagoya University, Furo, Chikusa, Nagoya, 464-8603, Japan

    Yoshinobu Baba

  8. Health Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, 761-0395, Japan

    Yoshinobu Baba

  9. Institute for Molecular Science, National Institutes of Natural Sciences, Okazaki, 444-8585, Japan

    Yoshinobu Baba

Authors
  1. Laili Mahmoudian
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  2. Jonas Melin
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  3. Mohamad Reza Mohamadi
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  4. Keiko Yamada
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  5. Michio Ohta
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  6. Noritada Kaji
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  7. Manabu Tokeshi
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  8. Mats Nilsson
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  9. Yoshinobu Baba
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Corresponding author

Correspondence to Laili Mahmoudian.

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Mahmoudian, L., Melin, J., Mohamadi, M.R. et al. Microchip Electrophoresis for Specific Gene Detection of the Pathogenic Bacteria V. cholerae by Circle-to-Circle Amplification. ANAL. SCI. 24, 327–332 (2008). https://doi.org/10.2116/analsci.24.327

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  • DOI: https://doi.org/10.2116/analsci.24.327

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