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 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.
ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France. julie.fredonnet@toulouse.inra.fr.
Julie
J
Fredonnet
ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France. julie.foncy@laas.fr.|LISBP, Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, Toulouse F-31077, France. julie.foncy@laas.fr.
Julie
J
Foncy
Innospys SAS, 3, allée des Vignes, Carbonne 31390, France. jc-cau@innopsys.fr.
Jean-Christophe
JC
Cau
ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France. childerick.severac@itav.fr.
Childérick
C
Séverac
ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France. fran_jm@insa-toulouse.fr.|LISBP, Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, Toulouse F-31077, France. fran_jm@insa-toulouse.fr.|Dendris SAS, 335 Rue du Chêne Vert, Labège 31670, France. fran_jm@insa-toulouse.fr.
Jean Marie
JM
François
ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France. emmanuelle.trevisiol@laas.fr.|CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France. emmanuelle.trevisiol@laas.fr.|LAAS, Univ de Toulouse, F-31400 Toulouse, France. emmanuelle.trevisiol@laas.fr.
Emmanuelle
E
Trévisiol
automation; dedicated microarrays; hybridization; microcontact printing; multiplexing; soft lithography
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