A SIMPLIFIED PROCEDURE FOR SYNTHESIZING NUCLEOSIDE 1-THIOTRIPHOSPHATES - DATP(ALPHA)S, DGTP(ALPHA)S, UTP-ALPHA-S, AND DTTP(ALPHA)S

A procedure for synthesizing a nucleoside 1-thiotriphosphate in a sing le reaction vessel beginning with the nucleoside, PSCl3, and PPi is de scribed. Reaction of the dried nucleoside with PSCl3 in anhydrous trie thylphosphate is followed by addition of the tetrabutylammonium salt o f PPi. Addition of excess triethylamine precipitates the nucleotides. The crude product is dissolved in water, and the nucleoside 1-thiotrip hosphate is purified in 24% to 50% yield by chromatography. The method is applied to the synthesis of dATP(alpha)S, dGTP(alpha)S, UTPalphaS and dTTP(alpha)S. (C) 1994 Academic Press, Inc.