Open AccessUse of 1,2,4-Dithiazolidine-3,5-Dione (DtsNH) and 3-Ethoxy-1,2,4-Dithiazoline-5-One (EDITH) for Synthesis of Phosphorothioate-Containing Oligodeoxyribonucleotides
Author(s) -
Qinghong Xu,
Karin MusierForsyth,
Robert P. Hammer,
George Bárány
Publication year - 1996
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/24.9.1602
Subject(s) - reagent , biology , alkoxy group , combinatorial chemistry , sulfur , disulfide bond , acetonitrile , chromatography , biochemistry , organic chemistry , chemistry , alkyl
Previous methods for the preparation of phosphorothioate-containing oligodeoxyribonucleotides rely on the reaction of phosphite triesters with sulfurizing reagents such as tetraethylthiuram disulfide (TETD) and 3H-1,2-benzodithiol-3-one 1,1-dioxide (Beaucage reagent). However, these and other sulfurizing reagents suffer from several disadvantages, and there is great impetus for the development of improved methods for sulfur transfer that are fully compatible with standard automated DNA synthesis. The present report describes the use of 1,2,4-dithiazolidine-3,5-dione (DtsNH) and 3-ethoxy-1,2,4-dithiazoline-5-one (EDITH) as effective sulfurizing reagents that meet these needs. Both reagents are easily prepared, and are stable upon prolonged room temperature storage in acetonitrile solution. The reagents are used at low concentrations (0.05 M) and for short reaction times (30 s). The methodology has been proven for the automated synthesis on 0.2-1.0 micromol scales of oligodeoxyribonucleotides, of length 6-20 bases, containing the phosphorothioate substitution at either a single site or at all positions.