title: Cross-platform comparison of nucleic acid hybridization: toward quantitative reference standards.
abstract: Measuring interactions between biological molecules is vitally important to both basic and applied research as well as development of pharmaceuticals. Although a wide and growing range of techniques is available to measure various kinetic and thermodynamic properties of interacting biomolecules, it can be difficult to compare data across techniques of different laboratories and personnel or even across different instruments using the same technique. Here we evaluate relevant biological interactions based on complementary DNA and RNA oligonucleotides that could be used as reference standards for many experimental systems. We measured thermodynamics of duplex formation using isothermal titration calorimetry, differential scanning calorimetry, and ultraviolet-visible (UV-vis) monitored denaturation/renaturation. These standards can be used to validate results, compare data from disparate techniques, act as a teaching tool for laboratory classes, or potentially to calibrate instruments. The RNA and DNA standards have many attractive features, including low cost, high purity, easily measurable concentrations, and minimal handling concerns, making them ideal for use as a reference material.
authors.affiliation: The RNA Institute, University at Albany-SUNY, Albany, NY 12222, USA. Electronic address: khalvorsen@albany.edu.
authors.forename: Ken
authors.initials: K
authors.lastname: Halvorsen
authors.affiliation: The RNA Institute, University at Albany-SUNY, Albany, NY 12222, USA; Department of Biological Sciences, University at Albany-SUNY, Albany, NY 12222, USA.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Agris
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D009843:Oligoribonucleotides / Q000737:chemistry*; D012313:RNA / Q000032:analysis* / Q000737:chemistry; D012015:Reference Standards
chemical_list: D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D012313:RNA; D004247:DNA
keywords: Biomolecular interaction analysis; DNA; Differential scanning calorimetry; Isothermal titration calorimetry; Oligonucleotides; RNA