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Aubert, Y ; Bourgerie, S ; Meunier, L ; Mayer, R ; Roche, AC ; Monsigny, M ; Thuong, NT ; Asseline, U

Optimized synthesis of phosphorothioate oligodeoxyribonucleotides substituted with a 5’-protected thiol function and a 3 ’-amino group

Nucleic Acids Research 28 (3) 818-825

par Administrateur - publié le , mis à jour le

Abstract :

A new deprotection procedure enables a medium scale preparation of phosphodiester and phosphorothioate oligonucleotides substituted with a protected thiol function at their 5’-ends and an amino group at their 3’-ends in good yield (up to 72 OD units/mu mol for a 19mer phosphorothioate). Syntheses of 3’-amino-substituted oligonucleotides were carried out on a modified support. A linker containing the thioacetyl moiety was manually coupled in two steps by first adding its phosphoramidite derivative in the presence of tetrazole followed by either oxidation or sulfurization to afford the bis-derivatized oligonucleotide bound to the support. Deprotection was achieved by treating the fully protected oligonucleotide with a mixture of 2,2’-dithiodipyridine and concentrated aqueous ammonia in the presence of phenol and methanol.