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

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 &l...

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Detalles Bibliográficos
Autores principales: Moorcroft, Matthew J., Meuleman, Wouter R. A., Latham, Steven G., Nicholls, Thomas J., Egeland, Ryan D., Southern, Edwin M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1088307/
https://www.ncbi.nlm.nih.gov/pubmed/15870385
http://dx.doi.org/10.1093/nar/gni075
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author Moorcroft, Matthew J.
Meuleman, Wouter R. A.
Latham, Steven G.
Nicholls, Thomas J.
Egeland, Ryan D.
Southern, Edwin M.
author_facet Moorcroft, Matthew J.
Meuleman, Wouter R. A.
Latham, Steven G.
Nicholls, Thomas J.
Egeland, Ryan D.
Southern, Edwin M.
author_sort Moorcroft, Matthew J.
collection PubMed
description 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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spelling pubmed-10883072005-05-04 In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication Moorcroft, Matthew J. Meuleman, Wouter R. A. Latham, Steven G. Nicholls, Thomas J. Egeland, Ryan D. Southern, Edwin M. Nucleic Acids Res Methods Online 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. Oxford University Press 2005 2005-05-03 /pmc/articles/PMC1088307/ /pubmed/15870385 http://dx.doi.org/10.1093/nar/gni075 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Methods Online
Moorcroft, Matthew J.
Meuleman, Wouter R. A.
Latham, Steven G.
Nicholls, Thomas J.
Egeland, Ryan D.
Southern, Edwin M.
In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title_full In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title_fullStr In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title_full_unstemmed In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title_short In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
title_sort in situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1088307/
https://www.ncbi.nlm.nih.gov/pubmed/15870385
http://dx.doi.org/10.1093/nar/gni075
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