Ghosh S S, Musso G F
Nucleic Acids Res. 1987 Jul 10;15(13):5353-72. doi: 10.1093/nar/15.13.5353.
Coupling efficiencies for the covalent attachment of oligonucleotides (17-29 bases in length) to solid supports derivatized with alkyl-amino and -carboxylic functionalities have been determined. Attachment efficiencies of 60-80% were obtained for coated long-chain alkylamino controlled pore glass (CPG) supports. Similar efficiencies of immobilization were observed for carboxyl-bearing supports, which additionally exhibited lower levels of non-covalent binding. The extent of terminally linked oligonucleotide was determined to be 50-55% of the overall attachment in the carbodiimide-mediated coupling reaction of a 5'-aminohexyl phosphoramidate derivative of a 29-mer to Sephacryl carboxyl support. While lower overall efficiencies of attachment were obtained in the reaction with Sephacryl N-hydroxysuccinimide-activated carboxyl support, greater than 80% of this coupling results in end-attached oligonucleotides.
已测定了长度为17 - 29个碱基的寡核苷酸与用烷基氨基和羧基官能团衍生化的固体支持物共价连接的偶联效率。对于涂覆的长链烷基氨基可控孔径玻璃（CPG）支持物，获得了60 - 80%的连接效率。对于含羧基的支持物，观察到了类似的固定化效率，其还表现出较低水平的非共价结合。在将29聚体的5'-氨基己基亚磷酰胺衍生物与Sephacryl羧基支持物进行碳二亚胺介导的偶联反应中，末端连接的寡核苷酸的程度被确定为总连接量的50 - 55%。虽然在与Sephacryl N-羟基琥珀酰亚胺活化的羧基支持物的反应中获得的总连接效率较低，但这种偶联中超过80%的结果是末端连接的寡核苷酸。
