barnett-developmentofanautomateddna-2008.pdf (466.21 kB)
Development of an automated DNA purification module using a micro-fabricated pillar chip
journal contribution
posted on 2008-01-01, 00:00 authored by B Hindson, D Gutierrez, K Ness, A Makarewicz, T Metz, U Setlur, W Benett, J Loge, B Colston, Paul FrancisPaul Francis, Neil BarnettNeil Barnett, J DzenitisWe 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.
History
Journal
AnalystVolume
133Issue
2Pagination
248 - 255Publisher
Royal Society of ChemistryLocation
London, EnglandPublisher DOI
ISSN
0003-2654eISSN
1364-5528Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2008, Royal Society of ChemistryUsage metrics
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