Towards a real-time, label-free, diamond-based DNA sensorle

Vermeeren, V., Bijnens, N., Wenmackers, S., Daenen, M., Haenen, K., Williams, Oliver Aneurin ORCID: https://orcid.org/0000-0002-7210-3004, Ameloot, M., Vandeven, A., Wagner, P. and Michiels, L. 2007. Towards a real-time, label-free, diamond-based DNA sensorle. Langmuir 23 (26) , pp. 13193-13202. 10.1021/la702143d

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Abstract

Most challenging in the development of DNA sensors is the ability to distinguish between fully complementary target ssDNA (single-strand DNA) and 1-mismatch ssDNA. To deal with this problem, we performed impedance spectroscopy on DNA-functionalized nanocrystalline diamond (NCD) layers during hybridization and denaturation. In both reactions, a difference in behavior was observed for 1-mismatch target DNA and complementary target DNA in real-time. During real-time hybridization, a decrease of the impedance was observed at lower frequencies when the complementary target DNA was added, while the addition of 1-mismatch target ssDNA caused no significant change. Fitting these results to an electrical circuit demonstrates that this is correlated with a decrease of the depletion zone in the space charge region of the diamond. During real-time denaturation, differentiation between 1-mismatch and complementary target DNA was possible at higher frequencies. Denaturation of complementary DNA showed the longest exponential decay time of the impedance, while the decay time during 1-mismatch denaturation was the shortest. The real-time hybridization and denaturation experiments were carried out on different NCD samples in various buffer solutions at temperatures between 20 and 80 °C. It was revealed that the best results were obtained using a Microhyb hybridization buffer at 80 °C and 10× PCR buffer at 30 °C for hybridization and 0.1 M NaOH at temperatures above 40 °C for denaturation. We demonstrate that the combination of real-time hybridization spectra and real-time denaturation spectra yield important information on the type of target. This approach may allow a reliable identification of the mismatch sequence, which is the most biologically relevant.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Schools > Physics and Astronomy
Subjects:	Q Science > QC Physics
Publisher:	American Chemical Society
ISSN:	0743-7463
Last Modified:	21 Oct 2022 08:43
URI:	https://orca.cardiff.ac.uk/id/eprint/34171

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