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Nucleic Acids Research 2005 33(8):e75; doi:10.1093/nar/gni075
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Published online 3 May 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Methods Online

In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication

Matthew J. Moorcroft*, Wouter R. A. Meuleman, Steven G. Latham, Thomas J. Nicholls, Ryan D. Egeland and Edwin M. Southern1

Oxamer, Oxford Gene Technology Ltd Sandy Lane, Yarnton, Oxford OX5 1PF, UK 1Department of Biochemistry, University of Oxford South Parks Road, Oxford OX1 3QU, UK

*To whom correspondence should be addressed. Tel: +44 (0)1865 856340; Fax: +44 (0)1865 842116; Email: matt.moorcroft{at}ogt.co.uk

Received March 7, 2005. Revised April 12, 2005. Accepted April 12, 2005.

In this paper, we demonstrate in situ synthesis of oligonucleotide probes on poly(dimethylsiloxane) (PDMS) microchannels through use of conventional phosphoramidite chemistry. PDMS polymer was moulded into a series of microchannels using standard soft lithography (micro-moulding), with dimensions <100 µm. The surface of the PDMS was derivatized by exposure to ultraviolet/ozone followed by vapour phase deposition of glycidoxypropyltrimethoxysilane and reaction with poly(ethylene glycol) spacer, resulting in a reactive surface for oligonucleotide coupling. High, reproducible yields were achieved for both 6mer and 21mer probes as assessed by hybridization to fluorescent oligonucleotides. Oligonucleotide surface density was comparable with that obtained on glass substrates. These results suggest PDMS as a stable and flexible alternative to glass as a suitable substrate in the fabrication and synthesis of DNA microarrays.


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