Preparation of desoxycytidine-5'-[α-32P]triphosphate

Synthesis of [32P]dCMP was elaborated using either the Symons or the Biebricher method. Both methods are suitable but the yield depends on the quality (source, age) of the the Biebricher method is insensitive to the presence of water traces, however, it proceeds at a considerably higher temperature. The enzyme synthesis of [α-32P]dCTP from separated from E. coli and commercially available enzymes, such as nucleoside monophosphate kinase and nucleoside diphosphate kinase. The optimal synthesis conditions were found. Column chromatography on QAE Sephadex A 25 (HCO3)- and elution with NH4HCO3 of a concentration of 0.5 mol/l were found suitable for [α-32P]dCTP purification. (author)