Webster K R, Shamoo Y, Konigsberg W, Spicer E K
Yale University School of Medicine, New Haven, CT 06510-0824.
Biotechniques. 1991 Nov;11(5):658-61.
A procedure for large-scale purification of synthetic oligoribonucleotides has been developed that has significant advantages over gel purification techniques currently in use. Synthesis was performed using commercially available 2'-O-silylated ribonucleoside 3'-O-phosphoramidites, and coupling efficiencies were consistently greater than 97% for oligoribonucleotides up to 31 residues in length. Using C4 reverse-phase chromatography to remove material not deprotected by treatment with tetrabutylammonium fluoride, we have eliminated reactants in which the 2'-O-silyl group is only partly removed, thus ensuring a homogeneous population of oligoribonucleotide.
已开发出一种大规模纯化合成寡核糖核苷酸的方法,该方法相对于目前使用的凝胶纯化技术具有显著优势。合成使用市售的2'-O-硅烷基化核糖核苷3'-O-亚磷酰胺进行,对于长度达31个残基的寡核糖核苷酸,偶联效率始终大于97%。通过使用C4反相色谱去除未用四丁基氟化铵处理脱保护的物质,我们消除了其中2'-O-硅烷基仅部分去除的反应物,从而确保了寡核糖核苷酸群体的均一性。