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Rapid, sequence-specific detection of unpurified PCR amplicons via a reusable, electrochemical sensor.


ABSTRACT: We report an electrochemical method for the sequence-specific detection of unpurified amplification products of the gyrB gene of Salmonella typhimurium. Using an asymmetric PCR and the electrochemical E-DNA detection scheme, single-stranded amplicons were produced from as few as 90 gene copies and, without subsequent purification, rapidly identified. The detection is specific; the sensor does not respond when challenged with control oligonucleotides based on the gyrB genes of either Escherichia coli or various Shigella species. In contrast to existing sequence-specific optical- and capillary electrophoresis-based detection methods, the E-DNA sensor is fully electronic and requires neither cumbersome, expensive optics nor high voltage power supplies. Given these advantages, E-DNA sensors appear well suited for implementation in portable PCR microdevices directed at, for example, the rapid detection of pathogens.

SUBMITTER: Lai RY 

PROVIDER: S-EPMC1449638 | biostudies-literature | 2006 Mar

REPOSITORIES: biostudies-literature

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Rapid, sequence-specific detection of unpurified PCR amplicons via a reusable, electrochemical sensor.

Lai Rebecca Y RY   Lagally Eric T ET   Lee Sang-Ho SH   Soh H T HT   Plaxco Kevin W KW   Heeger Alan J AJ  

Proceedings of the National Academy of Sciences of the United States of America 20060303 11


We report an electrochemical method for the sequence-specific detection of unpurified amplification products of the gyrB gene of Salmonella typhimurium. Using an asymmetric PCR and the electrochemical E-DNA detection scheme, single-stranded amplicons were produced from as few as 90 gene copies and, without subsequent purification, rapidly identified. The detection is specific; the sensor does not respond when challenged with control oligonucleotides based on the gyrB genes of either Escherichia  ...[more]

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