High lane density slab-gel electrophoresis using micromachined instrumentation.
In this paper, micromachined pipette arrays (MPAs) and microcombs were studied as a means of enabling high lane density gel electrophoresis. The MPA provide a miniaturized format to interface sub-microliter volumes of samples between macroscale sample preparation formats and microscale biochemical analysis systems. The microcombs provide a means of creating sample loading wells in the gel material on the same center-to-center spacing as the MPAs. Together, the two micromachined instruments provide an alternative to current combs and pipetting technologies used for creating sample loading wells and sample delivery in gel electrophoresis systems. Using three designs for the microcomb-MPA pair, center-to-center spacings of 1.0 mm, 500 microm, and 250 microm are studied. The results demonstrate an approximate 10-fold increase in lane density and a 10-fold reduction in sample size from 5 microL to 500 pL. As a result, the number of theoretical plates has increased 2.5-fold, while system resolution has increased 1.5-fold over the conventional agarose gel systems. An examination of changes in resolution across the width of individual separation lanes in both systems revealed dependence in the case of the conventional gels and no dependence for the gels loaded with the micromachined instrumentation.
Papautsky
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Department of Electrical and Computer Engineering and Computer Science, University of Cincinnati, OH 45221, USA. ian.papautsky@uc.edu
Mohanty
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Weiss
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R
Frazier
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D001978:Bromphenol Blue; D004396:Coloring Agents; D004247:DNA / Q000032:analysis; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation*; D004867:Equipment Design; D008832:Microchemistry / Q000295:instrumentation*; D008904:Miniaturization; D013048:Specimen Handling
D004396:Coloring Agents; D001978:Bromphenol Blue; D004247:DNA