Kumar P, Gupta K C
Nucleic Acids Research Laboratory, Institute of Genomics and Integrative Biology (formerly Centre for Biochemical Technology), Mall Road, Delhi University Campus, Delhi-110 007, India.
Bioconjug Chem. 2003 May-Jun;14(3):507-12. doi: 10.1021/bc025646q.
A simple method has been devised to construct oligonucleotide array on a variety of surfaces using commonly available reagents and chemistry with good efficiency and accuracy. The method involves the generation of hydroxyl functionalities on glass, polypropylene, polyethylene, and commonly used surfaces for construction of oligonucleotide arrays followed by their activation with trifluoroethanesulfonyl chloride (tresyl chloride). The activated surface in the subsequent reaction is used to covalently immobilize oligonucleotides in regioselective fashion to create an oligonucleotide array. The surface bound tresyl sulfonate esters allow the immobilization of oligonucleotides specifically via their 3'- or 5'-end having mercaptohexyl- or aminohexyl functionalities. The constructed oligonucleotide arrays were successfully used to analyze oligonucleotides by hybridization technique.
已设计出一种简单方法,可使用常用试剂和化学方法在各种表面高效且准确地构建寡核苷酸阵列。该方法包括在玻璃、聚丙烯、聚乙烯及构建寡核苷酸阵列常用的表面上生成羟基官能团,随后用三氟乙烷磺酰氯(三氟甲磺酰氯)对其进行活化。在后续反应中,活化后的表面用于以区域选择性方式共价固定寡核苷酸,从而创建寡核苷酸阵列。表面结合的三氟甲磺酸酯允许通过具有巯基己基或氨基己基官能团的3'-或5'-末端特异性固定寡核苷酸。构建的寡核苷酸阵列已成功用于通过杂交技术分析寡核苷酸。