Synthesis of the 3′-thio-nucleosides and subsequent automated synthesis of oligodeoxynucleotides containing a 3′-S-phosphorothiolate linkage

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Abstract
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.