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Development of an automated DNA purification module using a micro-fabricated pillar chip

Hindson, Benjamin J., Gutierrez, Dora M., Ness, Kevin D., Makarewicz, Anthony J., Metz, Thomas R., Setlur, Ujwal S., Benett, William B., Loge, Jeffrey M., Colston, Bill W. Jr., Francis, Paul S., Barnett, Neil W. and Dzenitis, John M. 2008, Development of an automated DNA purification module using a micro-fabricated pillar chip, Analyst, vol. 133, no. 2, pp. 248-255.

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Title Development of an automated DNA purification module using a micro-fabricated pillar chip
Author(s) Hindson, Benjamin J.
Gutierrez, Dora M.
Ness, Kevin D.
Makarewicz, Anthony J.
Metz, Thomas R.
Setlur, Ujwal S.
Benett, William B.
Loge, Jeffrey M.
Colston, Bill W. Jr.
Francis, Paul S.
Barnett, Neil W.
Dzenitis, John M.
Journal name Analyst
Volume number 133
Issue number 2
Start page 248
End page 255
Publisher Royal Society of Chemistry
Place of publication London, England
Publication date 2008
ISSN 0003-2654
1364-5528
1744-1560
Summary We present a fully automated DNA purification module comprised of a micro-fabricated chip and sequential injection analysis system that is designed for use within autonomous instruments that continuously monitor the environment for the presence of biological threat agents. The chip has an elliptical flow channel containing a bed (3.5 × 3.5 mm) of silica-coated pillars with height, width and center-to-center spacing of 200, 15, and 30 µm, respectively, which provides a relatively large surface area (ca. 3 cm2) for DNA capture in the presence of chaotropic agents. We have characterized the effect of various fluidic parameters on extraction performance, including sample input volume, capture flow rate, and elution volume. The flow-through design made the pillar chip completely reusable; carryover was eliminated by flushing lines with sodium hypochlorite and deionized water between assays. A mass balance was conducted to determine the fate of input DNA not recovered in the eluent. The device was capable of purifying and recovering Bacillus anthracis genomic DNA (input masses from 0.32 to 320 pg) from spiked environmental aerosol samples, for subsequent analysis using polymerase chain reaction-based assays.
Language eng
Field of Research 030103 Flow Analysis
HERDC Research category C1 Refereed article in a scholarly journal
HERDC collection year 2008
Copyright notice ©2008, Royal Society of Chemistry
Persistent URL http://hdl.handle.net/10536/DRO/DU:30017240

Document type: Journal Article
Collection: School of Life and Environmental Sciences
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