Characterization of dynamic solid phase DNA extraction from blood with magnetically controlled silica beads

• Published in: Analyst (London) 2010-03, Vol.135 (3), p.531-537
• Main Authors: DUARTE, Gabriela R. M, PRICE, Carol W, LITTLEWOOD, Janice L, HAVERSTICK, Doris M, FERRANCE, Jerome P, CARRILHO, Emanuel, LANDERS, James P
• Format: Article
• Language: English
• Subjects: Analytical chemistry; beta-Globins - genetics; Chemistry; DNA - blood; DNA - isolation & purification; Exact sciences and technology; Forensic Genetics; Humans; Magnetics; Point-of-Care Systems; Silicon Dioxide - chemistry; Solid Phase Extraction - methods; Sonication
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/b918996c

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.