DeDionisio L, Raible A M, Gryaznov S M
Lynx Therapeutics Incorporated, Hayward, CA 94545, USA.
Electrophoresis. 1998 Jun;19(8-9):1265-9. doi: 10.1002/elps.1150190807.
Oligonucleotide N3'-->P5' phosphoramidates (3'-phosphoramidates) are DNA analogs that are presently under investigation as potential therapeutic agents. These compounds may also hold promise as a diagnostic tool. Here, we describe a rapid method for the analysis of single-stranded RNA fragments utilizing 3'-phosphoramidate oligonucleotides as probes in conjunction with capillary gel electrophoresis (CGE). 3'-Phosphoramidate 9-mers were mixed with complimentary RNA, and CGE was used to monitor duplex formation. Complimentary strands of RNA and 3'-phosphoramidate formed duplexes that gave unique relative mobilities based on an internal standard. The ability of CGE to discriminate between perfect duplexes and duplexes that contain a base mismatch was also investigated. However, the primary focus of this work was to determine CGE's ability to detect the presence of the 3'-phosphoramidates/RNA duplex under routine electrophoretic running conditions. Polyacrylamide gel electrophoresis analysis was utilized to verify duplex formation.
寡核苷酸N3'→P5'氨基磷酸酯（3'-氨基磷酸酯）是一类DNA类似物，目前正作为潜在治疗药物进行研究。这些化合物也有望成为一种诊断工具。在此，我们描述了一种利用3'-氨基磷酸酯寡核苷酸作为探针结合毛细管凝胶电泳（CGE）分析单链RNA片段的快速方法。将3'-氨基磷酸酯九聚体与互补RNA混合，并用CGE监测双链体的形成。RNA和3'-氨基磷酸酯的互补链形成双链体，基于内标给出独特的相对迁移率。还研究了CGE区分完美双链体和含有碱基错配的双链体的能力。然而，这项工作的主要重点是确定CGE在常规电泳运行条件下检测3'-氨基磷酸酯/RNA双链体存在的能力。利用聚丙烯酰胺凝胶电泳分析来验证双链体的形成。
