title: Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification.
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.
authors.affiliation: Department of Applied Chemistry, Graduates School of Engineering, Nagoya University, Furo, Chikusa, Nagoya, Japan. mahmoudian2@yahoo.com
authors.forename: Laili
authors.initials: L
authors.lastname: Mahmoudian
authors.forename: Jonas
authors.initials: J
authors.lastname: Melin
authors.forename: Mohamad Reza
authors.initials: MR
authors.lastname: Mohamadi
authors.forename: Keiko
authors.initials: K
authors.lastname: Yamada
authors.forename: Michio
authors.initials: M
authors.lastname: Ohta
authors.forename: Noritada
authors.initials: N
authors.lastname: Kaji
authors.forename: Manabu
authors.initials: M
authors.lastname: Tokeshi
authors.forename: Mats
authors.initials: M
authors.lastname: Nilsson
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Baba
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D004270:DNA, Circular / Q000737:chemistry*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014734:Vibrio cholerae / Q000737:chemistry* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D004270:DNA, Circular