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Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays

Microarrays are established research tools for genotyping, expression profiling, or molecular diagnostics in which DNA molecules are precisely addressed to the surface of a solid support. This study assesses the fabrication of low-density oligonucleotide arrays using an automated microcontact printi...

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Autores principales: Fredonnet, Julie, Foncy, Julie, Cau, Jean-Christophe, Séverac, Childérick, François, Jean Marie, Trévisiol, Emmanuelle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5197944/
https://www.ncbi.nlm.nih.gov/pubmed/27681742
http://dx.doi.org/10.3390/microarrays5040025
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author Fredonnet, Julie
Foncy, Julie
Cau, Jean-Christophe
Séverac, Childérick
François, Jean Marie
Trévisiol, Emmanuelle
author_facet Fredonnet, Julie
Foncy, Julie
Cau, Jean-Christophe
Séverac, Childérick
François, Jean Marie
Trévisiol, Emmanuelle
author_sort Fredonnet, Julie
collection PubMed
description Microarrays are established research tools for genotyping, expression profiling, or molecular diagnostics in which DNA molecules are precisely addressed to the surface of a solid support. This study assesses the fabrication of low-density oligonucleotide arrays using an automated microcontact printing device, the InnoStamp 40(®). This automate allows a multiplexed deposition of oligoprobes on a functionalized surface by the use of a MacroStamp(TM) bearing 64 individual pillars each mounted with 50 circular micropatterns (spots) of 160 µm diameter at 320 µm pitch. Reliability and reuse of the MacroStamp(TM) were shown to be fast and robust by a simple washing step in 96% ethanol. The low-density microarrays printed on either epoxysilane or dendrimer-functionalized slides (DendriSlides) showed excellent hybridization response with complementary sequences at unusual low probe and target concentrations, since the actual probe density immobilized by this technology was at least 10-fold lower than with the conventional mechanical spotting. In addition, we found a comparable hybridization response in terms of fluorescence intensity between spotted and printed oligoarrays with a 1 nM complementary target by using a 50-fold lower probe concentration to produce the oligoarrays by the microcontact printing method. Taken together, our results lend support to the potential development of this multiplexed microcontact printing technology employing soft lithography as an alternative, cost-competitive tool for fabrication of low-density DNA microarrays.
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spelling pubmed-51979442017-01-04 Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays Fredonnet, Julie Foncy, Julie Cau, Jean-Christophe Séverac, Childérick François, Jean Marie Trévisiol, Emmanuelle Microarrays (Basel) Article Microarrays are established research tools for genotyping, expression profiling, or molecular diagnostics in which DNA molecules are precisely addressed to the surface of a solid support. This study assesses the fabrication of low-density oligonucleotide arrays using an automated microcontact printing device, the InnoStamp 40(®). This automate allows a multiplexed deposition of oligoprobes on a functionalized surface by the use of a MacroStamp(TM) bearing 64 individual pillars each mounted with 50 circular micropatterns (spots) of 160 µm diameter at 320 µm pitch. Reliability and reuse of the MacroStamp(TM) were shown to be fast and robust by a simple washing step in 96% ethanol. The low-density microarrays printed on either epoxysilane or dendrimer-functionalized slides (DendriSlides) showed excellent hybridization response with complementary sequences at unusual low probe and target concentrations, since the actual probe density immobilized by this technology was at least 10-fold lower than with the conventional mechanical spotting. In addition, we found a comparable hybridization response in terms of fluorescence intensity between spotted and printed oligoarrays with a 1 nM complementary target by using a 50-fold lower probe concentration to produce the oligoarrays by the microcontact printing method. Taken together, our results lend support to the potential development of this multiplexed microcontact printing technology employing soft lithography as an alternative, cost-competitive tool for fabrication of low-density DNA microarrays. MDPI 2016-09-26 /pmc/articles/PMC5197944/ /pubmed/27681742 http://dx.doi.org/10.3390/microarrays5040025 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fredonnet, Julie
Foncy, Julie
Cau, Jean-Christophe
Séverac, Childérick
François, Jean Marie
Trévisiol, Emmanuelle
Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title_full Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title_fullStr Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title_full_unstemmed Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title_short Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays
title_sort automated and multiplexed soft lithography for the production of low-density dna microarrays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5197944/
https://www.ncbi.nlm.nih.gov/pubmed/27681742
http://dx.doi.org/10.3390/microarrays5040025
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