Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification. |
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| Authors: | Laili Mahmoudian Jonas Melin Mohamad Reza Mohamadi Keiko Yamada Michio Ohta Noritada Kaji Manabu Tokeshi Mats Nilsson Yoshinobu Baba |
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| Institution: | Department of Applied Chemistry, Graduates School of Engineering, Nagoya University, Furo, Chikusa, Nagoya, Japan. mahmoudian2@yahoo.com |
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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 degrees 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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