title: Light emitting diode, photodiode-based fluorescence detection system for DNA analysis with microchip electrophoresis.
abstract: Electrophoretic separation of fluorescently end-labeled DNA after a PCR serves as a gold standard in genetic diagnostics. Because of their size and cost, instruments for this type of analysis have had limited market uptake, particularly for point-of-care applications. This might be changed through a higher level of system integration and lower instrument costs that can be realized through the use of LEDs for excitation and photodiodes for detection--if they provide sufficient sensitivity. Here, we demonstrate an optimized microchip electrophoresis instrument using polymeric fluidic chips with fluorescence detection of end-labeled DNA with a LOD of 0.15 nM of Alexa Fluor 532. This represents orders of magnitude improvement over previously reported instruments of this type. We demonstrate the system with an electrophoretic separation of two PCR products and their respective primers. We believe that this is the first LED-induced fluorescence microchip electrophoresis system with photodiode-based detection that could be used for standard applications of PCR and electrophoresis.
authors.affiliation: Department of Electrical and Computer Engineering, University of Waterloo, ON, Canada.
authors.forename: Gordon H
authors.initials: GH
authors.lastname: Hall
authors.affiliation: Department of Biology, University of Waterloo, ON, Canada.|Waterloo Institute of Nanotechnology, University of Waterloo, ON, Canada.
authors.forename: D Moira
authors.initials: DM
authors.lastname: Glerum
authors.affiliation: Department of Electrical and Computer Engineering, University of Waterloo, ON, Canada.|Waterloo Institute of Nanotechnology, University of Waterloo, ON, Canada.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Backhouse
mesh_terms: D004247:DNA / Q000032:analysis*; D004581:Electronics; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D057230:Limit of Detection
chemical_list: D004247:DNA
keywords: Genetic analysis; Light emitting diode induced fluorescence detection; Microchip electrophoresis; Microfluidics; Photodiode detection