Analysis of Modified Nucleosides Using Glass Microfluidic Devices (5 pages)
By D.L. Morris, Jr., C.P. Thiede, A.M. Huehls, C.F. Sweeny, A.Z. Siahmakoun, Rose-Hulman Institute of Technology
Conference: 2006 2006 SEM Annual Conference & Exposition on Experimental and Applied Mechanics
Nucleoside derivatives are important markers for diagnosing and/or monitoring serious diseases and clinical conditions. These markers include the ubiquitous oxidative DNA damage marker 8-hydroxy-2-deoxyguanosine (8-OH-dG). With the exclusion of ionizing radiation, oxidative DNA damage is associated with the metal-dependent decomposition of H2O2 and subsequent generation of reactive oxygen species (ROS). However, the relationship between site specific binding of metal ions to DNA and RNA and the extent of oxidative damage arising from this binding is not well characterized. We probe these interactions by monitoring 8-OH-dG production from nucleosides and nucleotides using glass microfluidic devices with interferometric backscattering detection (IBSD). While laser-induced fluorescence (LIF) is typically employed as a detection scheme for separations performed on glass microfluidic devices, IBSD is a reasonable alternative to LIF for detecting analytes that do not fluoresce, as is the case for nucleosides and their derivatives. IBSD responds to refractive index changes, making it a universal detection method, and reported detection limits rival those of LIF. The detection characteristics of IBSD and the superior separation characteristics of electrophoretic separations on microfluidic devices allow us to study and characterize metal ion interactions with nucleosides and nucleotides and how these interactions affect 8-OH-dG production.