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Electric field-induced concentration and capture of DNA onto microtips
Authors:Dinesh Kalyanasundaram  Shinnosuke Inoue  Jong-Hoon Kim  Hyun-Boo Lee  Zenko Kawabata  Woon-Hong Yeo  Gerard A Cangelosi  Kieseok Oh  Dayong Gao  Kyong-Hoon Lee  Jae-Hyun Chung
Affiliation:1. Department of Mechanical Engineering, University of Washington, Seattle, WA, 98195, USA
2. Seattle Biomedical Research Institute, 307 Westlake Ave N, Suite 500, Seattle, WA, 98109, USA
3. NanoFacture, Inc., P.O. Box 52651, Bellevue, WA, 98015, USA
Abstract:Simple, high-yield concentration of DNA is important for high-throughput genetic analysis and disease diagnosis. Glass-based microfilters are popular but the process requires centrifugation steps with cumbersome chemical processes. As an alternative, a concentration method using an electric field has been explored previously, but with limited efficiency. In this paper, electric field-induced concentration and capture of DNA are studied by using high-aspect-ratio microtips coated with a gold layer. The microtips are immersed longitudinally into a solution of 100???L containing ??-phage DNA. After DNA concentration using an electric field, the microtips are withdrawn from the solution. Under AC- and biased AC fields, DNA is concentrated by electrophoresis (EP), dielectrophoresis (DEP), and electroosmotic flow (EOF). To reduce capillary effects in the withdrawal process, the microtips are coated with positively charged poly-l-lysine (PLL). The pattern of captured DNA is analyzed by fluorescence microscopy. DEP attracts DNA molecules at the edges of microtips, where the highest gradient of electric field exists. EP attracts DNA onto the surface of microtips following the vectors of an electric field. EOF generates vortexes that deliver DNA onto microtips. Using this method, 85% of DNA is captured on the PLL-coated microtips after three sequential captures. The concentration mechanism can potentially facilitate rapid and simple preparation of DNA for downstream analysis.
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