Synthesis of the 3′-thio-nucleosides and subsequent automated synthesis of oligodeoxynucleotides containing a 3′-S-phosphorothiolate linkage
Synthesis of the 3′-thio-nucleosides and subsequent automated synthesis of oligodeoxynucleotides containing a 3′-S-phosphorothiolate linkage
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DOI：
10.1038/nprot.2007.451
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Cosstick, Richard
中科院分区：
文献类型：
--
作者：
Gaynor, James W.;Bentley, Joanne;Cosstick, Richard
含有3'- s -磷硫酸酯（3'-PS）键的寡脱氧核苷酸已成为探测酶催化的DNA裂解过程的有用工具。本方案描述了由胸腺嘧啶和2'-脱氧胞苷衍生的磷硫酰胺单体的合成，以及它们在合成含有3'-PS键的寡脱氧核苷酸的全自动程序中的应用。5'-protected-3'-amidites的合成在2周内完成，DNA的合成和纯化需要1周。
Oligodeoxynucleotides containing 3'-S-phosphorothiolate (3'-PS) linkages have become useful tools for probing enzyme-catalyzed cleavage processes in DNA. This protocol describes the synthesis of the phosphorothioamidite monomers derived from thymidine and 2'-deoxycytidine, and their application to a fully automated procedure for synthesising oligodeoxynucleotides containing 3'-PS linkages. The synthesis of the 5'-protected-3'-amidites is achievable in 2 weeks with the DNA synthesis and purification taking another 1 week.