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Development of a bi-functional silica monolith for electro-osmotic pumping and DNA clean-up/extraction using gel-supported reagents in a microfluidic device

Oakley, Jennifer A.; Shaw, Kirsty J.; Docker, Peter T.; Dyer, Charlotte E.; Greenman, John; Greenway, Gillian M.; Haswell, Stephen J.

Authors

Jennifer A. Oakley

Kirsty J. Shaw

Peter T. Docker

Charlotte E. Dyer C.E.Dyer@hull.ac.uk

Gillian M. Greenway G.M.Greenway@hull.ac.uk

Stephen J. Haswell



Abstract

A silica monolith used to support both electro-osmotic pumping (EOP) and the extraction/elution of DNA coupled with gel-supported reagents is described. The benefits of the combined EOP extraction/elution system were illustrated by combining DNA extraction and gene amplification using the polymerase chain reaction (PCR) process. All the reagents necessary for both processes were supported within pre-loaded gels that allow the reagents to be stored at 4 degrees C for up to four weeks in the microfluidic device. When carrying out an analysis the crude sample only needed to be hydrodynamically introduced into the device which was connected to an external computer controlled power supply via platinum wire electrodes. DNA was extracted with 65% efficiency after loading lysed cells onto a silica monolith. Ethanol contained within an agarose gel matrix was then used to wash unwanted debris away from the sample by EOP (100 V cm(-1) for 5 min). The retained DNA was subsequently eluted from the monolith by water contained in a second agarose gel, again by EOP using an electric field of 100 V cm(-1) for 5 min, and transferred into the PCR reagent containing gel. The eluted DNA in solution was successfully amplified by PCR, confirming that the concept of a complete self-contained microfluidic device could be realised for DNA sample clean up and amplification, using a simple pumping and on-chip reagent storage methodology.

Journal Article Type Article
Publication Date 2009
Journal LAB ON A CHIP
Print ISSN 1473-0197
Electronic ISSN 1473-0189
Publisher Royal Society of Chemistry
Peer Reviewed Peer Reviewed
Volume 9
Issue 11
Pages 1596-1600
APA6 Citation Oakley, J. A., Shaw, K. J., Docker, P. T., Dyer, C. E., Greenman, J., Greenway, G. M., & Haswell, S. J. (2009). Development of a bi-functional silica monolith for electro-osmotic pumping and DNA clean-up/extraction using gel-supported reagents in a microfluidic device. Lab on a chip, 9(11), 1596-1600. https://doi.org/10.1039/b820553a
DOI https://doi.org/10.1039/b820553a
Keywords Biochemistry; Bioengineering; General Chemistry; Biomedical Engineering
Publisher URL https://pubs.rsc.org/en/Content/ArticleLanding/2009/LC/b820553a#!divAbstract
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