Microchip-Based Macroporous Silica Sol−Gel Monolith for Efficient Isolation of DNA from Clinical Samples
- 8 July 2006
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 78 (16), 5704-5710
- https://doi.org/10.1021/ac060390t
Abstract
Effective microchip extraction of deoxyribonucleic acid (DNA) from crude biological matrixes has been demonstrated using silica beads or hybrid phases composed of beads and sol−gel. However, the use of monolithic sol−gels alone for extraction of human genomic DNA has been more difficult to define. Here we describe, for the first time, the successful use of monolithic tetramethyl orthosilicate-based sol−gels for effective micro-solid-phase extraction (μSPE) of DNA in a glass microchip format. A functional monolithic silica phase with micrometer-scale pores in the silica matrix resulted from addition of poly(ethylene glycol), a poragen, to the precursor mixture. This allowed a monolithic sol−gel bed to be established in a microchip channel that provided large surface area for DNA extraction with little flow-induced back pressure. DNA extraction efficiencies for simple systems (λ-phage DNA) were ∼85%, while efficiencies for the reproducible extraction of human genomic DNA from complex biological matrixes (human blood) were ∼70%. Blockage of the sol−gel pores by components in the lysed blood was observed in repeat extraction on a single device as a decrease in the extraction efficiency. The developed μSPE protocol was further evaluated to show applicability to clinical samples and bacterial cultures, through extraction of PCR-amplifiable DNA.Keywords
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