Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification

Laili Mahmoudian; Jonas Melin; Mohamad Reza Mohamadi; Keiko Yamada; Michio Ohta; Noritada Kaji; Manabu Tokeshi; Mats Nilsson; Yoshinobu Baba

doi:10.2116/analsci.24.327

Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification

Laili Mahmoudian, Jonas Melin, Mohamad Reza Mohamadi, Keiko Yamada, Michio Ohta, Noritada Kaji, Manabu Tokeshi, Mats Nilsson, Yoshinobu Baba

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

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. 2008

Original language	English
Pages (from-to)	327-332
Number of pages	6
Journal	analytical sciences
Volume	24
Issue number	3
DOIs	https://doi.org/10.2116/analsci.24.327
Publication status	Published - Mar 2008
Externally published	Yes

All Science Journal Classification (ASJC) codes

Analytical Chemistry

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10.2116/analsci.24.327

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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°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. 2008",

author = "Laili Mahmoudian and Jonas Melin and Mohamadi, {Mohamad Reza} and Keiko Yamada and Michio Ohta and Noritada Kaji and Manabu Tokeshi and Mats Nilsson and Yoshinobu Baba",

year = "2008",

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journal = "Analytical Sciences",

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T1 - Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification

AU - Mahmoudian, Laili

AU - Melin, Jonas

AU - Mohamadi, Mohamad Reza

AU - Yamada, Keiko

AU - Ohta, Michio

AU - Kaji, Noritada

AU - Tokeshi, Manabu

AU - Nilsson, Mats

AU - Baba, Yoshinobu

PY - 2008/3

Y1 - 2008/3

N2 - 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. 2008

AB - 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. 2008

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JO - Analytical Sciences

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Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification

Abstract

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