DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice |
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| Authors: | K.R. Jackson J.C. Borba M. Meija D.L. Mills D.M. Haverstick K.E. Olson R. Aranda G.T. Garner E. Carrilho J.P. Landers |
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| Affiliation: | 1. Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA;2. Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, VA 22904, USA;3. Department of Pathology, University of Virginia, Charlottesville, VA 22904, USA;4. Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos-SP, Brazil;5. TeGrex Technologies, Sperryville, VA 22740, USA;6. Office of the Chief Scientist, Defense Forensic Science Center, N 31st Street, Atlanta, GA 30297, USA |
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| Abstract: | We report the development of a disposable polyester toner centrifugal device for semi-automated, dynamic solid phase DNA extraction (dSPE) from whole blood samples. The integration of a novel adhesive and hydrophobic valving with a simple and low cost microfabrication method allowed for sequential addition of reagents without the need for external equipment for fluid flow control. The spin-dSPE method yielded an average extraction efficiency of ∼45% from 0.6 μL of whole blood. The device performed single sample extractions or accommodate up to four samples for simultaneous DNA extraction, with PCR-readiness DNA confirmed by effective amplification of a β-globin gene. The purity of the DNA was challenged by a multiplex amplification with 16 targeted amplification sites. Successful multiplexed amplification could routinely be obtained using the purified DNA collected post an on-chip extraction, with the results comparable to those obtained with commercial DNA extraction methods. This proof-of-principle work represents a significant step towards a fully-automated low cost DNA extraction device. |
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| Keywords: | DNA purification dSPE PeT device Polyethylene terephthalate |
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