Automated and multiplexed soft lithography for the production of low-density DNA microarrays

Julie Fredonnet (1) , Julie Foncy (1, 2) , Jean Christophe Cau (3) , Childérick Séverac (1) , Jean François (1, 2, 4) , Emmanuelle Trévisiol (1, 5)

1 ITAV - Institut des Technologies Avancées en sciences du Vivant
2 LISBP - Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés
3 Innospys SAS
4 Dendris SAS
5 LAAS-ELIA - Équipe Ingénierie pour les sciences du vivant

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.

Mots clés: automation; dedicated microarrays; hybridization; microcontact printing; multiplexing; soft lithography

Mots clés: hybridation adn; génotypage

Domaines: Sciences du Vivant [q-bio]

Julie Fredonnet, Julie Foncy, Jean Christophe Cau, Childérick Séverac, Jean François, et al.. Automated and multiplexed soft lithography for the production of low-density DNA microarrays. Microarrays, 2016, 5 (4), pp.25. ⟨10.3390/microarrays5040025⟩. ⟨hal-01601906⟩