title: Real-time monitoring of DNA hybridization and melting processes using a fiber optic sensor.
abstract: In this paper a fiber optic surface plasmon resonance (FO-SPR) sensor was used to analyze the melting process of DNA linked to silica nanoparticles. Real-time monitoring of a DNA melting process has rarely been studied using surface plasmon resonance (SPR), since most commercial SPR setups do not allow for dynamic and accurate temperature control above 50 °C. The FO-SPR sensor platform, with silica nanobead signal amplification, allows sensing inside a standard PCR thermocycler, which makes high resolution DNA melting curve analysis possible. This innovative combination was used to characterize the hybridization and melting events between DNA immobilized on the sensor surface and DNA probes on silica nanoparticles. At optimized hybridization conditions complementary DNA strands of different lengths could be distinguished. While the real-time FO-SPR analysis of DNA hybridization did not result in significant variances, the analysis of DNA melting determined the exact length of overlap and the matching Gibbs energy.
authors.affiliation: Department of Biosystems, Division Mechatronics, Biostatistics and Sensors, KULeuven, Leuven, Belgium.
authors.forename: Filip
authors.initials: F
authors.lastname: Delport
authors.forename: Jeroen
authors.initials: J
authors.lastname: Pollet
authors.forename: Kris
authors.initials: K
authors.lastname: Janssen
authors.forename: Bert
authors.initials: B
authors.lastname: Verbruggen
authors.forename: Karel
authors.initials: K
authors.lastname: Knez
authors.forename: Dragana
authors.initials: D
authors.lastname: Spasic
authors.forename: Jeroen
authors.initials: J
authors.lastname: Lammertyn
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry; D004867:Equipment Design; D053758:Nanoparticles / Q000737:chemistry*; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D012822:Silicon Dioxide / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D015342:DNA Probes; D012822:Silicon Dioxide; D004247:DNA