用于高效杂交平台的DNA探针在（DMA-NAS-MAPS）共聚物涂层基板上的反应性微接触印刷

Reactive Microcontact Printing of DNA Probes on (DMA-NAS-MAPS) Copolymer-Coated Substrates for Efficient Hybridization Platforms.

作者信息

Castagna Rossella, Bertucci Alessandro, Prasetyanto Eko Adi, Monticelli Marco, Conca Dario Valter, Massetti Matteo, Sharma Parikshit Pratim, Damin Francesco, Chiari Marcella, De Cola Luisa, Bertacco Riccardo

机构信息

Dipartimento di Elettronica Informazione e Bioingegneria, Politecnico di Milano , Via G. Colombo 81, 20133, Milano, Italy.

Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg , 8 allée Gaspard Monge, 67000, Strasbourg, France.

出版信息

Langmuir. 2016 Apr 5;32(13):3308-13. doi: 10.1021/acs.langmuir.5b04669. Epub 2016 Mar 22.

DOI:10.1021/acs.langmuir.5b04669

PMID:26972953

Abstract

High-performing hybridization platforms fabricated by reactive microcontact printing of DNA probes are presented. Multishaped PDMS molds are used to covalently bind oligonucleotides over a functional copolymer (DMA-NAS-MAPS) surface. Printed structures with minimum width of about 1.5 μm, spaced by 10 μm, are demonstrated, with edge corrugation lower than 300 nm. The quantification of the immobilized surface probes via fluorescence imaging gives a remarkable concentration of 3.3 × 10(3) oligonucleotides/μm(2), almost totally active when used as probes in DNA-DNA hybridization assays. Indeed, fluorescence and atomic force microscopy show a 95% efficiency in target binding and uniform DNA hybridization over printed areas.

摘要

介绍了通过DNA探针的反应性微接触印刷制备的高性能杂交平台。使用多形状的聚二甲基硅氧烷（PDMS）模具在功能性共聚物（DMA-NAS-MAPS）表面共价结合寡核苷酸。展示了最小宽度约为1.5μm、间距为10μm的印刷结构，其边缘波纹低于300nm。通过荧光成像对固定在表面的探针进行定量分析，得到了3.3×10³寡核苷酸/μm²的显著浓度，在DNA-DNA杂交试验中用作探针时几乎完全具有活性。实际上，荧光和原子力显微镜显示在印刷区域的靶标结合效率为95%，且DNA杂交均匀。