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In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication

Glass has become the standard substrate for the preparation of DNA arrays. Typically, glass is modified using silane chemistries to provide an appropriate functional group for nucleic acid synthesis or oligonucleotide immobilization. We have found substantial issues with the stability of these surfa...

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Autores principales: Phillips, Margaret F., Lockett, Matthew R., Rodesch, Matthew J., Shortreed, Michael R., Cerrina, Franco, Smith, Lloyd M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2248760/
https://www.ncbi.nlm.nih.gov/pubmed/18084027
http://dx.doi.org/10.1093/nar/gkm1103
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author Phillips, Margaret F.
Lockett, Matthew R.
Rodesch, Matthew J.
Shortreed, Michael R.
Cerrina, Franco
Smith, Lloyd M.
author_facet Phillips, Margaret F.
Lockett, Matthew R.
Rodesch, Matthew J.
Shortreed, Michael R.
Cerrina, Franco
Smith, Lloyd M.
author_sort Phillips, Margaret F.
collection PubMed
description Glass has become the standard substrate for the preparation of DNA arrays. Typically, glass is modified using silane chemistries to provide an appropriate functional group for nucleic acid synthesis or oligonucleotide immobilization. We have found substantial issues with the stability of these surfaces as manifested in the unwanted release of oligomers from the surface when incubated in aqueous buffers at moderate temperatures. To address this issue, we have explored the use of carbon-based substrates. Here, we demonstrate in situ synthesis of oligonucleotide probes on carbon-based substrates using light-directed photolithographic phosphoramidite chemistry and evaluate the stabilities of the resultant DNA arrays compared to those fabricated on silanized glass slides. DNA arrays on carbon-based substrates are substantially more stable than arrays prepared on glass. This superior stability enables the use of high-density DNA arrays for applications involving high temperatures, basic conditions, or where serial hybridization and dehybridization is desired.
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spelling pubmed-22487602008-02-21 In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication Phillips, Margaret F. Lockett, Matthew R. Rodesch, Matthew J. Shortreed, Michael R. Cerrina, Franco Smith, Lloyd M. Nucleic Acids Res Methods Online Glass has become the standard substrate for the preparation of DNA arrays. Typically, glass is modified using silane chemistries to provide an appropriate functional group for nucleic acid synthesis or oligonucleotide immobilization. We have found substantial issues with the stability of these surfaces as manifested in the unwanted release of oligomers from the surface when incubated in aqueous buffers at moderate temperatures. To address this issue, we have explored the use of carbon-based substrates. Here, we demonstrate in situ synthesis of oligonucleotide probes on carbon-based substrates using light-directed photolithographic phosphoramidite chemistry and evaluate the stabilities of the resultant DNA arrays compared to those fabricated on silanized glass slides. DNA arrays on carbon-based substrates are substantially more stable than arrays prepared on glass. This superior stability enables the use of high-density DNA arrays for applications involving high temperatures, basic conditions, or where serial hybridization and dehybridization is desired. Oxford University Press 2008-01 2007-12-15 /pmc/articles/PMC2248760/ /pubmed/18084027 http://dx.doi.org/10.1093/nar/gkm1103 Text en © 2007 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Phillips, Margaret F.
Lockett, Matthew R.
Rodesch, Matthew J.
Shortreed, Michael R.
Cerrina, Franco
Smith, Lloyd M.
In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title_full In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title_fullStr In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title_full_unstemmed In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title_short In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
title_sort in situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2248760/
https://www.ncbi.nlm.nih.gov/pubmed/18084027
http://dx.doi.org/10.1093/nar/gkm1103
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