A general method for the synthesis of 5'-monophosphates of DNA fragments via phosphotriester intermediates.

A new and attractive phosphorylation procedure which allows the introduction, via phosphotriester intermediates, of 5'-phosphate functions of DNA fragments is described. The method is based on the activation of bifunctional phosphorylating agents with 1-hydroxybenzotriazole. The approach will be exemplified by the synthesis of pACGC using four different 5'-phosphotriester intermediates.