Screening and electrochemical detection of an antibiotic producing gene in bacteria on an integrated microchip
- Authors
- Han, Dawoon; Chand, Rohit; Shin, Ik-Soo; Kim, Yong-Sang
- Issue Date
- 2013
- Publisher
- ROYAL SOC CHEMISTRY
- Citation
- ANALYTICAL METHODS, v.5, no.23, pp.6814 - 6820
- Journal Title
- ANALYTICAL METHODS
- Volume
- 5
- Number
- 23
- Start Page
- 6814
- End Page
- 6820
- URI
- http://scholarworks.bwise.kr/ssu/handle/2018.sw.ssu/12298
- DOI
- 10.1039/c3ay41321g
- ISSN
- 1759-9660
- Abstract
- We demonstrate the electrochemical determination of a secondary metabolite producing gene on an integrated microfluidic chip. The fabricated microchip was assembled with a continuous channel for polymerase chain reaction (PCR) and an electrochemical detector in order to achieve rapid and sensitive determination of the valC gene. valC is a gene responsible for producing antibiotic validamycin A in Streptomyces hygroscopicus. Biotin-conjugated primers amplified the valC gene. After the PCR, a DNA amplicon was analyzed in the electrochemical cell containing a streptavidin functionalized Au working electrode. The guanosine present in the DNA amplicon released electrons upon electrochemical oxidation at 0.93 V and the peak current linearly increased with the concentration of the captured DNA amplicon. The fabricated chip successfully amplified and detected the valC gene as low as 30 pg mu L-1 resulting in a sensitive, portable and integrated DNA analysis for a secondary metabolite.
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