Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification.
Anal Sci
; 24(3): 327-32, 2008 Mar.
Article
em En
| MEDLINE
| ID: mdl-18332538
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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Base de dados:
MEDLINE
Assunto principal:
Vibrio cholerae
/
DNA Bacteriano
/
DNA Circular
/
Técnicas de Amplificação de Ácido Nucleico
/
Eletroforese em Microchip
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article