Microchip-based purification of DNA from biological samples.
A microchip solid-phase extraction method for purification of DNA from biological samples, such as blood, is demonstrated. Silica beads were packed into glass microchips and the beads immobilized with sol-gel to provide a stable and reproducible solid phase onto which DNA could be adsorbed. Optimization of the DNA loading conditions established a higher DNA recovery at pH 6.1 than 7.6. This lower pH also allowed for the flow rate to be increased, resulting in a decrease in extraction time from 25 min to less than 15 min. Using this procedure, template genomic DNA from human whole blood was purified on the microchip platform with the only sample preparation being mixing of the blood with load buffer prior to loading on the microchip device. Comparison between the microchip SPE (microchipSPE) procedure and a commercial microcentrifuge method showed comparable amounts of PCR-amplifiable DNA could be isolated from cultures of Salmonella typhimurium. The greatest potential of the microchipSPE device was illustrated by purifying DNA from spores from the vaccine strain of Bacillus anthracis, where eventual integration of SPE, PCR, and separation on a single microdevice could potentially enable complete detection of the infectious agent in less than 30 min.
Breadmore
Michael C
MC
Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA.
Wolfe
Kelley A
KA
Arcibal
Imee G
IG
Leung
Wayne K
WK
Dickson
Dana
D
Giordano
Braden C
BC
Power
Mary E
ME
Ferrance
Jerome P
JP
Feldman
Sanford H
SH
Norris
Pamela M
PM
Landers
James P
JP
D000327:Adsorption; D001769:Blood; D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D006801:Humans; D008832:Microchemistry / Q000295:instrumentation; D008863:Microspheres; D012822:Silicon Dioxide
D004269:DNA, Bacterial; D012822:Silicon Dioxide; D004247:DNA