DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice

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

doi:10.1016/j.aca.2016.06.036

DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice

Anal Chim Acta. 2016 Sep 21:937:1-10. doi: 10.1016/j.aca.2016.06.036. Epub 2016 Jul 6.

Authors

K R Jackson¹, J C Borba², M Meija³, D L Mills⁴, D M Haverstick⁵, K E Olson⁶, R Aranda⁶, G T Garner³, E Carrilho², J P Landers⁷

Affiliations

¹ Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
² Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos-SP, Brazil.
³ Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, VA 22904, USA.
⁴ TeGrex Technologies, Sperryville, VA 22740, USA.
⁵ Department of Pathology, University of Virginia, Charlottesville, VA 22904, USA.
⁶ Office of the Chief Scientist, Defense Forensic Science Center, N 31st Street, Atlanta, GA 30297, USA.
⁷ 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.

PMID: 27590539
DOI: 10.1016/j.aca.2016.06.036

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.

Keywords: DNA purification; PeT device; Polyethylene terephthalate; dSPE.

MeSH terms

DNA / chemistry
DNA / genetics
DNA / isolation & purification*
Equipment Design
Hydrophobic and Hydrophilic Interactions
Lab-On-A-Chip Devices*
Magnetic Fields
Polyethylene Terephthalates / chemistry*
Polymerase Chain Reaction
Rotation*
Solid Phase Extraction / instrumentation*

Substances

Polyethylene Terephthalates
DNA