Characterization of dynamic solid phase DNA extraction from blood with magnetically controlled silica beads.
A novel solid phase extraction technique is described where DNA is bound and eluted from magnetic silica beads in a manner where efficiency is dependent on the magnetic manipulation of the beads and not on the flow of solution through a packed bed. The utility of this technique in the isolation of reasonably pure, PCR-amplifiable DNA from complex samples is shown by isolating DNA from whole human blood, and subsequently amplifying a fragment of the beta-globin gene. By effectively controlling the movement of the solid phase in the presence of a static sample, the issues associated with reproducibly packing a solid phase in a microchannel and maintaining consistent flow rates are eliminated. The technique described here is rapid, simple, and efficient, allowing for recovery of more than 60% of DNA from 0.6 microL of blood at a concentration which is suitable for PCR amplification. In addition, the technique presented here requires inexpensive, common laboratory equipment, making it easily adopted for both clinical point-of-care applications and on-site forensic sample analysis.
Duarte
Gabriela R M
GR
Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, Virginia 22904, USA.
Price
Carol W
CW
Littlewood
Janice L
JL
Haverstick
Doris M
DM
Ferrance
Jerome P
JP
Carrilho
Emanuel
E
Landers
James P
JP
D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D053803:Forensic Genetics; D006801:Humans; D008280:Magnetics; D019095:Point-of-Care Systems; D012822:Silicon Dioxide / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods*; D013010:Sonication; D055544:beta-Globins / Q000235:genetics
D055544:beta-Globins; D012822:Silicon Dioxide; D004247:DNA