Ellington A, Pollard J D
University of Texas, Austin, Texas, USA.
Curr Protoc Nucleic Acid Chem. 2001 May;Appendix 3:Appendix 3C. doi: 10.1002/0471142700.nca03cs00.
Modern nucleic acid synthesizers utilize phosphite triester chemistries that employ stable phosphoramidite monomers to build a growing polymer. These robust reactions allow easy generation of specific oligodeoxyribo- and oligoribonucleotides with a variety of labels, modified linkages, and nonstandard bases. Strategies are given for the maximization of synthetic yield, the generation of sequences containing site-specific modifications, and the isolation of synthetic oligonucleotides. Protocols describe monitoring the progress of synthesis via the trityl assay and methods for deprotection.
现代核酸合成仪利用亚磷酸三酯化学方法,该方法采用稳定的亚磷酰胺单体来构建不断增长的聚合物。这些稳健的反应能够轻松生成带有各种标记、修饰连接和非标准碱基的特定寡脱氧核糖核苷酸和寡核糖核苷酸。文中给出了使合成产率最大化、生成含有位点特异性修饰的序列以及分离合成寡核苷酸的策略。方案描述了通过三苯甲基测定法监测合成进程以及脱保护方法。