title: DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice.
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.
authors: Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
authors: K R
authors: KR
authors: Jackson
authors: Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos-SP, Brazil.
authors: J C
authors: JC
authors: Borba
authors: Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, VA 22904, USA.
authors: M
authors: M
authors: Meija
authors: TeGrex Technologies, Sperryville, VA 22740, USA.
authors: D L
authors: DL
authors: Mills
authors: Department of Pathology, University of Virginia, Charlottesville, VA 22904, USA.
authors: D M
authors: DM
authors: Haverstick
authors: Office of the Chief Scientist, Defense Forensic Science Center, N 31st Street, Atlanta, GA 30297, USA.
authors: K E
authors: KE
authors: Olson
authors: Office of the Chief Scientist, Defense Forensic Science Center, N 31st Street, Atlanta, GA 30297, USA.
authors: R
authors: R
authors: Aranda
authors: Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, VA 22904, USA.
authors: G T
authors: GT
authors: Garner
authors: Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos-SP, Brazil.
authors: E
authors: E
authors: Carrilho
authors: Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA; Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, VA 22904, USA; Department of Pathology, University of Virginia, Charlottesville, VA 22904, USA. Electronic address: landers@virginia.edu.
authors: J P
authors: JP
authors: Landers
mesh_terms: D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004867:Equipment Design; D057927:Hydrophobic and Hydrophilic Interactions; D056656:Lab-On-A-Chip Devices; D060526:Magnetic Fields; D011093:Polyethylene Terephthalates / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D012399:Rotation; D052616:Solid Phase Extraction / Q000295:instrumentation*
chemical_list: D011093:Polyethylene Terephthalates; D004247:DNA
keywords: DNA purification; PeT device; Polyethylene terephthalate; dSPE