An integrated microfluidic device for DNA purification and PCR amplification of STR fragments.
This work presents the integration of DNA extraction from complex samples and PCR amplification of STR fragments in a valveless, glass microdevice, using commercially available kits and instrumentation. DNA extraction was performed using a microchannel packed with a silica solid phase and a standard syringe pump as a single pressure source driving the extraction process, followed by integrated, online microchip amplification of STR fragments in a total volume of 1.2 microL. Reported characteristics important to this work include the capacity of the device for purification of DNA from a complex biological sample (whole blood) and the timing of DNA elution from the silica solid phase for successful downstream PCR amplification by placement the microdevice into a conventional thermocycler. Potential application of this microdevice to forensic genetic analysis was demonstrated through the preliminary extraction of DNA from semen, followed by an integrated, multiplexed, on-chip amplification that yielded detectable STR amplicons. By utilizing conventional laboratory equipment, the device presented exploits the benefits of microfluidic systems without complex control systems.
Bienvenue
Joan M
JM
Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA. joan.bienvenue@lmco.com
Legendre
Lindsay A
LA
Ferrance
Jerome P
JP
Landers
James P
JP
D004247:DNA / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D006801:Humans; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D016133:Polymerase Chain Reaction; D012661:Semen / Q000737:chemistry; D012822:Silicon Dioxide; D020080:Tandem Repeat Sequences
D012822:Silicon Dioxide; D004247:DNA