title: A valveless microfluidic device for integrated solid phase extraction and polymerase chain reaction for short tandem repeat (STR) analysis.
abstract: A valveless microdevice has been developed for the integration of solid phase extraction (SPE) and polymerase chain reaction (PCR) on a single chip for the short tandem repeat (STR) analysis of DNA from a biological sample. The device consists of two domains--a SPE domain filled with silica beads as a solid phase and a PCR domain with an ~500 nL reaction chamber. DNA from buccal swabs was purified and amplified using the integrated device and a full STR profile (16 loci) resulted. The 16 loci Identifiler® multiplex amplification was performed using a non-contact infrared (IR)-mediated PCR system built in-house, after syringe-driven SPE, providing an ~80-fold and 2.2-fold reduction in sample and reagent volumes consumed, respectively, as well as an ~5-fold reduction in the overall analysis time in comparison to conventional analysis. Results indicate that the SPE-PCR system can be used for many applications requiring genetic analysis, and the future addition of microchip electrophoresis (ME) to the system would allow for the complete processing of biological samples for forensic STR analysis on a single microdevice.
authors.affiliation: Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, VA 22904, USA.
authors.forename: Kristin A
authors.initials: KA
authors.lastname: Hagan
authors.forename: Carmen R
authors.initials: CR
authors.lastname: Reedy
authors.forename: Joan M
authors.initials: JM
authors.lastname: Bienvenue
authors.forename: Alison H
authors.initials: AH
authors.lastname: Dewald
authors.forename: James P
authors.initials: JP
authors.lastname: Landers
mesh_terms: D004247:DNA / Q000302:isolation & purification; D046209:Electrophoresis, Microchip; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D018895:Microsatellite Repeats; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012822:Silicon Dioxide / Q000737:chemistry; D052616:Solid Phase Extraction / Q000379:methods*
chemical_list: D007202:Indicators and Reagents; D012822:Silicon Dioxide; D004247:DNA