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Title: | Rapid assessment of the stability of DNA duplexes by impedimetric real-time monitoring of chemically induced denaturation | Authors: | VAN GRINSVEN, Bart VANDEN BON, Natalie GRIETEN, Lars MURIB, Mohammed Sharif JANSSENS, Stoffel HAENEN, Ken Schneider, E. Ingebrandt, S. Schöning, M.J. VERMEEREN, Veronique AMELOOT, Marcel MICHIELS, Luc THOELEN, Ronald DE CEUNINCK, Ward WAGNER, Patrick |
Issue Date: | 2011 | Source: | LAB ON A CHIP, 2011(9), p. 1656-1663 | Abstract: | In this article, we report on the electronic monitoring of DNA denaturation by NaOH using electrochemical impedance spectroscopy in combination with fluorescence imaging as a reference technique. The probe DNA consisting of a 36-mer fragment was covalently immobilized on nanocrystalline-diamond electrodes and hybridized with different types of 29-mer target DNA (complementary, single-nucleotide defects at two different positions, and a non-complementary random sequence). The mathematical separation of the impedimetric signals into the time constant for NaOH exposure and the intrinsic denaturation-time constants gives clear evidence that the denaturation times reflect the intrinsic stability of the DNA duplexes. The intrinsic time constants correlate with calculated DNA-melting temperatures. The impedimetric method requires minimal instrumentation, is label-free and fast with a typical time scale of minutes and is highly reproducible. The sensor electrodes can be used repetitively. These elements suggest that the monitoring of chemically induced denaturation at room temperature is an interesting approach to measure DNA duplex stability as an alternative to thermal denaturation at elevated temperatures, used in DNA-melting experiments and single nucleotide polymorphism (SNP) analysis. | Document URI: | http://hdl.handle.net/1942/13164 | ISSN: | 1473-0197 | e-ISSN: | 1473-0189 | DOI: | 10.1039/C1LC20027E | ISI #: | 000289363800012 | Rights: | This journal is © The Royal Society of Chemistry 2011 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2012 |
Appears in Collections: | Research publications |
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