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


Autoria(s): DUARTE, Gabriela R. M.; PRICE, Carol W.; LITTLEWOOD, Janice L.; HAVERSTICK, Doris M.; FERRANCE, Jerome P.; CARRILHO, Emanuel; LANDERS, James P.
Contribuinte(s)

UNIVERSIDADE DE SÃO PAULO

Data(s)

19/04/2012

19/04/2012

2010

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.

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2005/04473-4]

Identificador

ANALYST, v.135, n.3, p.531-537, 2010

0003-2654

http://producao.usp.br/handle/BDPI/16853

10.1039/b918996c

http://dx.doi.org/10.1039/b918996c

Idioma(s)

eng

Publicador

ROYAL SOC CHEMISTRY

Relação

Analyst

Direitos

closedAccess

Copyright ROYAL SOC CHEMISTRY

Palavras-Chave #CHEMICAL-ANALYSIS SYSTEMS #NUCLEIC-ACIDS #PURIFICATION #MICROCHIP #MONOLITH #CAPTURE #AGAROSE #SAMPLES #Chemistry, Analytical
Tipo

article

original article

publishedVersion