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Development of a real-world direct interface for integrated DNA extraction and amplification in a microfluidic device

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

Authors

Domino A. Joyce

Kirsty J. Shaw

Peter T. Docker

Charlotte E. Dyer

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

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

Stephen J. Haswell



Abstract

Integrated DNA extraction and amplification have been carried out in a microfluidic device using electro-osmotic pumping (EOP) for fluidic control. All the necessary reagents for performing both DNA extraction and polymerase chain reaction (PCR) amplification were pre-loaded into the microfluidic device following encapsulation in agarose gel. Buccal cells were collected using OmniSwabs [Whatman™, UK] and manually added to a chaotropic binding/lysis solution pre-loaded into the microfluidic device. The released DNA was then adsorbed onto a silica monolith contained within the DNA extraction chamber and the microfluidic device sealed using polymer electrodes. The washing and elution steps for DNA extraction were carried out using EOP, resulting in transfer of the eluted DNA into the PCR chamber. Thermal cycling, achieved using a Peltier element, resulted in amplification of the Amelogenin locus as confirmed using conventional capillary gel electrophoresis. It was demonstrated that the PCR reagents could be stored in the microfluidic device for at least 8 weeks at 4 °C with no significant loss of activity. Such methodology lends itself to the production of 'ready-to-use' microfluidic devices containing all the necessary reagents for sample processing, with many obvious applications in forensics and clinical medicine. © 2011 The Royal Society of Chemistry.

Journal Article Type Article
Publication Date 2011
Journal LAB ON A CHIP
Print ISSN 1473-0197
Electronic ISSN 1473-0189
Publisher Royal Society of Chemistry
Peer Reviewed Peer Reviewed
Volume 11
Issue 3
Pages 443-448
APA6 Citation Shaw, K. J., Joyce, D., Docker, P. T., Dyer, C. E., Greenway, G. M., Greenman, J., & Haswell, S. J. (2011). Development of a real-world direct interface for integrated DNA extraction and amplification in a microfluidic device. Lab on a chip, 11(3), 443-448. https://doi.org/10.1039/c0lc00346h
DOI https://doi.org/10.1039/c0lc00346h
Keywords Biochemistry; Bioengineering; General Chemistry; Biomedical Engineering
Publisher URL http://pubs.rsc.org/en/Content/ArticleLanding/2011/LC/C0LC00346H#!divAbstract
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