RRC ID 17396
Author Shiraishi H, Itoh T, Hayashi H, Takagi K, Sakane M, Mori T, Wang J.
Title Electrochemical detection of E. coli 16S rDNA sequence using air-plasma-activated fullerene-impregnated screen printed electrodes.
Journal Bioelectrochemistry
Abstract A new DNA modified electrode for the electrochemical detection of 16S rDNA extracted from Escherichia coli (JCM1649) is proposed. The electrodes were fabricated by screen printing a fullerene-impregnated carbon ink onto a poly(methylmethacrylate) substrate and immobilizing a probe DNA on the surface after activating the electrode with air plasma. The results indicated a dramatic improvement in the surface coverage of the immobilized probe DNA, and of the reduction peak of the redox indicator (Co(phen)(3)(3+)) due to the incorporation of fullerene. By immobilizing the probe onto the fullerene-impregnated screen-printed electrodes, the PCR product of the 16S rDNA extracted from E. coli was directly detected without any pretreatment. A well defined signal difference was observed between the perfectly matching oligonucleotide and the mismatching one, and it was possible to detect the target at the modified electrode. This method enabled us to clearly detect the two base mismatches in the ca. 1500-bases long 16S rDNA sequence.
Volume 70(2)
Pages 481-7
Published 2007-5-1
DOI 10.1016/j.bioelechem.2006.07.011
PII S1567-5394(06)00101-0
PMID 16962387
MeSH Air Base Pair Mismatch DNA, Bacterial / chemistry DNA, Bacterial / genetics* DNA, Ribosomal / chemistry DNA, Ribosomal / genetics* Electrochemistry / instrumentation Electrochemistry / methods* Electrodes Escherichia coli / genetics* Fullerenes / chemistry Hot Temperature In Situ Hybridization / instrumentation In Situ Hybridization / methods* RNA, Ribosomal, 16S / chemistry RNA, Ribosomal, 16S / genetics* Sequence Analysis, DNA / methods* Surface Properties
IF 4.722
Times Cited 39
WOS Category BIOLOGY BIOPHYSICS ELECTROCHEMISTRY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
General Microbes JCM 1649