Direct loading of polymer matrices in plastic microchips for rapid DNA analysis: a comparative study.
We report the design and performance validation of microfluidic separation technologies for human identification using a disposable plastic device suitable for integration into an automated rapid DNA analysis system. A fabrication process for a 15-cm long hot-embossed plastic microfluidic devices with a smooth semielliptical cross section out of cyclic olefin copolymer is presented. We propose a mixed polymer solution of 95% w/v hydroxyethylcellulose and 5% w/v polyvinylpyrrolidone for a final polymer concentration of 2.5 or 3.0% to be used as coating and sieving matrix for DNA separation. This formulation allows preparing the microchip without pretreatment in a single-loading step and provides high-resolution separation (≈1.2 bp for fragments <200 bp), which is superior to existing commercial matrices under the same conditions. The hot-embossed device performance is characterized and compared to injection-molded devices made out of cyclic olefin copolymer based on their respective injector geometry, channel shape, and surface charges. Each device design is assessed by fluorescence videomicroscopy to evaluate the formation of injection plugs, then by comparing electropherograms for the separation of a DNA size standard relevant to human identification.
Center for Applied Nanobioscience and Medicine, The University of Arizona College of Medicine, Phoenix, AZ 85004, USA. churth@email.arizona.edu
Cedric
C
Hurth
Jian
J
Gu
Maurice
M
Aboud
Matthew D
MD
Estes
Alan R
AR
Nordquist
Bruce
B
McCord
Frederic
F
Zenhausern
D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D006801:Humans; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D011205:Povidone / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D004247:DNA; D011205:Povidone