title: Rapid assessment of the stability of DNA duplexes by impedimetric real-time monitoring of chemically induced denaturation.
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.
authors: Hasselt University, Institute for Materials Research, Wetenschapspark 1, B-3590, Diepenbeek, Belgium. bart.vangrinsven@uhasselt.be
authors: B
authors: B
authors: van Grinsven
authors: N
authors: N
authors: Vanden Bon
authors: L
authors: L
authors: Grieten
authors: M
authors: M
authors: Murib
authors: S D
authors: SD
authors: Janssens
authors: K
authors: K
authors: Haenen
authors: E
authors: E
authors: Schneider
authors: S
authors: S
authors: Ingebrandt
authors: M J
authors: MJ
authors: Schöning
authors: V
authors: V
authors: Vermeeren
authors: M
authors: M
authors: Ameloot
authors: L
authors: L
authors: Michiels
authors: R
authors: R
authors: Thoelen
authors: W
authors: W
authors: De Ceuninck
authors: P
authors: P
authors: Wagner
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D058266:Dielectric Spectroscopy / Q000379:methods*; D018613:Microscopy, Confocal; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012972:Sodium Hydroxide / Q000737:chemistry; D044366:Transition Temperature
chemical_list: D015342:DNA Probes; D012972:Sodium Hydroxide; D004247:DNA