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

Autor(es)

Duarte, Gabriela R. M. ^{[1, 2, 3]} ; Price, Carol W. ^[3] ; Littlewood, Janice L. ^[3] ; Haverstick, Doris M. ^[4] ; Ferrance, Jerome P. ^[3] ; Carrilho, Emanuel ^{[1, 2]} ; Landers, James P. ^{[3, 5, 4]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Quim Sao Carlos, BR-13566590 Sao Carlos, SP - Brazil

^[2] Inst Nacl Ciencia & Tecnol Bioanalit, BR-13083970 Campinas, SP - Brazil

^[3] Univ Virginia, Dept Chem, Charlottesville, VA 22904 - USA

^[4] Univ Virginia, Hlth Sci Ctr, Dept Pathol, Charlottesville, VA 22908 - USA

^[5] Univ Virginia, Dept Mech Engn, Charlottesville, VA 22904 - USA

• Tipo de documento: Artigo Científico
• Fonte: ANALYST; v. 135, n. 3, p. 531-537, 2010.
• Citações Web of Science: 30

Resumo

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 mu L 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. (AU)

• Processo FAPESP: 05/04473-4 - Análises genéticas em sistemas nanofabricados
• Beneficiário: Gabriela Rodrigues Mendes Duarte
• Modalidade de apoio: Bolsas no Brasil - Doutorado