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.
Similar content being viewed by others
References
International Human Genome Sequencing Consortium, Nature, 2004, 431, 931.
A. C. Syvanen, Nat. Genet., 2005, 37(Suppl), S5.
M. Nilsson, F. Dahl, C. Larsson, M. Gullberg, and J. Stenberg, Trends Biotechnol., 2006, 24, 83.
H. Tian, L. C. Brody, and J. P. Landers, Genome. Res., 2000, 10, 1403.
Y. Hataoka, L. Zhang, Y. Mori, N. Tomita, T. Notomi, and Y. Baba, Anal. Chem., 2004, 76, 3689.
Y. Hataoka, L. Zhang, T. Yukimasa, and Y. Baba, Anal. Sci., 2005, 21, 53.
A. Manz, N. Graber, and H. M. Widmer, Sens. Actuators, B, 1990, 1, 244.
F. Xu, M. Jabasini, and Y. Baba, Electrophoresis, 2002, 23, 3608.
M. Nilsson, H. Malmgren, M. Samiotaki, M. Kwiatkowski, B. P. Chowdhary, and U. Landegren, Science, 1994, 265, 2085.
F. Dahl, J. Baner, M. Gullberg, M. Mendel-Hartvig, and U. Landegren, and M. Nilsson, Proc. Natl. Acad. Sci. U. S. A., 2006, 101, 4548.
J. Jarvius, J. Melin, J. Goransson, J. Stenberg, S. Fredriksson, C. Gonzalez-Rey, S. Bertilsson, and M. Nilsson, Nat. Methods, 2006, 3, 725.
L. Mahmoudian, J. Melin, M. R. Mohamadi, K. Yamada, M. Ohta, N. Kaji, M. Tokeshi, M. Nilsson, and Y. Baba, Chem. Lett., 2007, 36, 396.
Y. Baba, N. Ishimura, K. Samata, and M. Tsuhako, J. Chromatogr., A, 1993, 653, 329.
F. Xu, M. Jabasini, and Y. Baba, Electrophoresis, 2005, 26, 3013.
J. Stenberg, M. Nilsson, and U. Landegren, BMC Bioinformatics, 2005, 6, 299.
L. Blanco, A. Bernad, J. Lazare, G. Martin, C. Garmendia, and M. Salas, J. Biol. Chem., 1989, 264, 8935.
M. R. Mohamadi, M. Kataoka, L. Mahmoudian, M. Jabasini, Y. Shinohara, and Y. Baba, Chromatographia, 2005, 61, 339.
J. Banner, A. Isaksson, E. Waldenstrom, J. Jarvius, U. Landegren, and M. Nilsson, Nucleic Acids Res., 2003, 31, el03.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2116/analsci.24.327