DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice
Creators
- 1. Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States)
- 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 (United States)
- 4. TeGrex Technologies, Sperryville, VA 22740 (United States)
- 5. Department of Pathology, University of Virginia, Charlottesville, VA 22904 (United States)
- 6. Office of the Chief Scientist, Defense Forensic Science Center, N 31st Street, Atlanta, GA 30297 (United States)
Description
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. - Highlights: • dSPE design on centrifugal PeT device with a unique mixing strategy was proposed. • Increased fluidic control with novel adhesive tape valves on a PeT device. • Multiplexed spin-dSPE device to run up to 4 samples simultaneously. • Demonstrated strong singleplexed and multiplexed amplification following chip dSPE.
Availability note (English)
Available from http://dx.doi.org/10.1016/j.aca.2016.06.036Additional details
Identifiers
- DOI
- 10.1016/j.aca.2016.06.036;
- PII
- S0003-2670(16)30799-1;
Publishing Information
- Journal Title
- Analytica Chimica Acta
- Journal Volume
- 937
- Journal Page Range
- p. 1-10
- ISSN
- 0003-2670
- CODEN
- ACACAM
INIS
- Country of Publication
- Netherlands
- Country of Input or Organization
- International Atomic Energy Agency (IAEA)
- INIS RN
- 48096966
- Subject category
- S37: INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY;
- Resource subtype / Literary indicator
- Numerical Data
- Descriptors DEI
- AMPLIFICATION; DNA; EXPERIMENTAL DATA; EXTRACTION; FLUID FLOW; POLYESTERS; POSITRON COMPUTED TOMOGRAPHY
- Descriptors DEC
- COMPUTERIZED TOMOGRAPHY; DATA; DIAGNOSTIC TECHNIQUES; EMISSION COMPUTED TOMOGRAPHY; ESTERS; INFORMATION; NUCLEIC ACIDS; NUMERICAL DATA; ORGANIC COMPOUNDS; ORGANIC POLYMERS; POLYMERS; SEPARATION PROCESSES; TOMOGRAPHY
Optional Information
- Copyright
- Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.