Electrochemical quantification of DNA amplicons via the detection of non-hybridised guanine bases on low-density electrode arrays
| Autor: | Olivier Y.F. Henry, Daniel Latta, J.L. Acero Sanchez, Ciara K. O'Sullivan |
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| Rok vydání: | 2009 |
| Předmět: |
Guanine
Biomedical Engineering Biophysics Analytical chemistry Electrochemistry Sensitivity and Specificity chemistry.chemical_compound In Situ Hybridization Oligonucleotide Array Sequence Analysis Reproducibility of Results DNA Equipment Design General Medicine Amplicon Combinatorial chemistry Equipment Failure Analysis Microelectrode chemistry Electrode Microelectrodes Nucleic Acid Amplification Techniques Biosensor Methylene blue Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 24:2064-2070 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2008.10.021 |
| Popis: | A new strategy for the electrochemical detection and signal amplification of DNA at gold electrodes is described. Current methodologies for DNA biosensing based on the electrochemical detection of electroactive base-specific labels such as methylene blue (MB) suffer from lengthy incubation and washing steps. Addressing these limitations, we report a novel approach for the electrochemical quantification of surface hybrid, using the control gene LTA, 107 bases long, as a model target. An array of 15 gold electrodes was used to detect the formation of hybridised duplex following interaction of non-hybridised guanine bases with MB present in solution. Upon hybridisation the number of free guanines present at the electrode surface increased from 8 to 25 due to guanine bases present in the target sequence which did not participate in hybridisation and remained free to interact directly with methylene blue. This increase in free guanines consequently concentrated MB directly at the electrode surface. We found that the MB signal recorded for 100 nM of the complementary LTA was typically 2.14 times higher than that of the non-hybridised state. Very low cross-reactivity ( |
| Databáze: | OpenAIRE |
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