title: The effect of temperature on electrochemically driven denaturation monitored by SERS.
abstract: Scanning the electrochemical potential negative results in the gradual denaturation of dsDNA immobilised at a nanostructure gold electrode, the DNA melting is monitored by SERS. We demonstrate the effect of the experimental temperature on the electrochemically driven melting (E-melting) by carrying out experiments between 10 and 28 °C using two DNA duplexes (20 and 21 base pairs). Significant temperature dependence for both the melting potentials, Em, and the steepness of the melting curves was found over the range 10 to 18 °C. Above 18 °C the results were found to be independent of temperature. The relative temperature insensitivity of the melting potentials above 18 °C is advantageous for the application of the electrochemically driven melting technique because precise temperature control is not necessary for measurements that are carried out around room temperature. Conversely temperature dependence below 18 °C offers a way to improve discrimination for highly similar DNA sequences.
authors.affiliation: Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK.
authors.forename: Evanthia
authors.initials: E
authors.lastname: Papadopoulou
authors.affiliation: Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK.
authors.forename: Marta
authors.initials: M
authors.lastname: Meneghello
authors.affiliation: Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford OX1 3TA, UK.
authors.forename: Pietro
authors.initials: P
authors.lastname: Marafini
authors.affiliation: Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Johnson
authors.affiliation: Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford OX1 3TA, UK.
authors.forename: Tom
authors.initials: T
authors.lastname: Brown
authors.affiliation: Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK. Electronic address: P.N.Bartlett@soton.ac.uk.
authors.forename: Philip N
authors.initials: PN
authors.lastname: Bartlett
mesh_terms: D004247:DNA / Q000737:chemistry; D061925:Immobilized Nucleic Acids / Q000737:chemistry; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013696:Temperature
chemical_list: D061925:Immobilized Nucleic Acids; D009692:Nucleic Acid Heteroduplexes; D004247:DNA
keywords: Biosensor; DNA stability; E-melting; Electrochemically driven melting; SERS; Temperature