Assembly fabrication of linkers on glass surface and their effect on DNA synthesis and hybridization  

玻片表面手臂分子的组装及对寡核苷酸原位合成与杂交的影响(英文)

作　　者：沈佳尧[1] 肖鹏峰[1] 侯鹏[1] 祭美菊[1] 孙啸[1] 何农跃[1] 

机构地区：[1]东南大学生物科学与医学工程系,吴健雄实验室,南京210096

出　　处：《Journal of Southeast University(English Edition)》2003年第2期149-154,共6页东南大学学报（英文版）

基　　金：TheNationalNaturalScienceFoundationofChina(6 0 0 710 0 1)

摘　　要：Linkers were assembled on a glass surface based on the hydrolysis and condensation of 3-glycidoxy￣propyltrimethoxysilane (GPS). After the assembly of GPS, four approaches were tried to open the ending epoxide group of GPS or to further elongate the linkers. The effect of these approaches on DNA in situ synthesis and hybridization was investigated. For the spacing of the synthesis initiation sites, the wettability of the support and the length of the linking group that attaches the initiation site to the surface have direct influences on the yield of coupling reactions and the subsequent hybridization events. X-ray photoelectron spectroscopy (XPS) and mean contact angles of deionized water of the above slides were measured to assess the linker's characteristics in each procedure. It was proved that the glass slides were successfully modified and became excellent supports for the oligonucleotides synthesis. In addition, it proved best for the in situ oligonucleotides synthesis that a glass slide was in turn treated with ethylenediamine, glutaradehyde, ethanolamine and sodium borohydride solution at ambient temperature after silanized with GPS.以 3缩水甘油环氧丙基三甲氧基硅烷对玻璃基片表面进行硅烷化后 ,用下列 4种方法组装手臂分子 :①盐酸直接处理 ;②先用聚六乙二醇 ,然后用盐酸处理 ;③用乙二胺、戊二醛、乙醇胺和硼氢化钠分别处理 ;④用聚六乙二醇、乙二胺、戊二醛、乙醇胺和硼氢化钠分别处理 .用XPS(X-ray photoelectron spectroscopy)和测定接触角的方法对上述组装进行了表征 ,并用直接偶联荧光单体及合成 2 0mer寡核苷酸与带荧光的互补探针杂交的方法对上述手臂分子的合成效率及杂交效率进行了考察 .实验表明方法③组装的手臂分子得到的结果优于其他 3种方法 ,证明了手臂分子的空间效应、亲水性等性质对寡核苷酸合成和杂交存在影响 .

关 键 词：LINKER assembly fabrication in situ oligonucleotide synthesis HYBRIDIZATION 

分 类 号：O641.3[理学—物理化学] O629.74[理学—化学]