title: Rapid nucleic acid melting analyses using a microfabricated electrochemical platform.
abstract: Microfabrication methods have been used to fabricate a new microscale platform that integrates thermal control and multi-electrode components to enable rapid, temperature-dependent electrochemical measurements on small-volume fluid samples. A wide range of biochemical phenomena can be characterized with the device, for example, when monitoring interactions at the working electrode between probe and target species which include an electroactive moiety. Employing square wave voltammetry, we have demonstrated the utility and reproducibility of the microplatform in melting studies on full-match, single-mismatch, and double-mismatch DNA structures of relevance to single-nucleotide polymorphism (SNP) discrimination. As shown, the small size of the reported device, low volume for the samples it can interrogate (∼10 μL), individual addressing of platform components and fast localized heating (settling times ∼5 s) combine to allow for efficient sample analyses. In addition, a straight-forward route exists, involving replication into array formats and integration with microfluidics, for extending the technology toward eventual high throughput work on drug discovery and medical diagnostics.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20740, USA.
authors.forename: Zuliang
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20740, USA. Electronic address: hsintim@umd.edu.
authors.forename: Herman O
authors.initials: HO
authors.lastname: Sintim
authors.affiliation: Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. Electronic address: stephen.semancik@nist.gov.
authors.forename: Steve
authors.initials: S
authors.lastname: Semancik
mesh_terms: D020137:Base Pair Mismatch; D004247:DNA / Q000737:chemistry*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation; D055616:Microtechnology; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D044366:Transition Temperature
chemical_list: D009841:Oligonucleotides; D004247:DNA
keywords: DNA melting curves; Microfabricated electrochemical platform; Nucleic acid self-assembly; Rapid temperature control; Single nucleotide polymorphism