Centro de Reconocimiento Molecular y Desarrollo Tecnológico, Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
Bioconjug Chem. 2012 Oct 17;23(10):2121-8. doi: 10.1021/bc300333a. Epub 2012 Sep 28.
Selective covalent immobilization of thiolated oligonucleotides onto an epoxy-functionalized silicon-substrate can be achieved via light radiation (365 nm). Following this approach, thiol-modified oligonucleotide probes were covalently attached as microarrays, reaching an immobilization density of 2.5 pmol·cm(-2), with a yield of 53%. The developed method presents the advantages of spatially controlled probe anchoring (by means of using a photomask), direct attachment without using cross-linkers, and short irradiation times (10 min). Hybridization efficiencies up to 65%, with full complementary strands, were reached. The approach was evaluated by scoring single nucleotide polymorphisms with a discrimination ratio around 15. Moreover, sensitive and selective detection of bacterial Escherichia coli was demonstrated.
通过光辐射（365nm）可以实现将巯基修饰的寡核苷酸选择性地共价固定在环氧功能化的硅衬底上。采用这种方法，将硫醇修饰的寡核苷酸探针作为微阵列进行共价固定，固定密度达到 2.5pmol·cm(-2)，产率为 53%。该方法具有探针固定的空间可控性（通过使用光掩模）、无需使用交联剂的直接固定以及短的辐射时间（10 分钟）等优点。达到了高达 65%的杂交效率，完全互补链。该方法通过评分单核苷酸多态性，其区分率约为 15。此外，还证明了对细菌大肠杆菌的敏感和选择性检测。
