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A rapid quantitative method for measuring UTP, UDP, and UMP in the picomole range.
Authors:C P Cheung  R J Suhadolnik
Institution:Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140 USA
Abstract:A procedure for the determination of picomole amounts of uracil nucleotides is described. The key reaction is the condensation of UTP and 14C]glucose 1-phosphate catalyzed by uridine 5′-diphosphoglucose pyrophosphorylase yielding UDP-14C]glucose. The product is determined by selective adsorption onto charcoal in the presence of 0.8 m Trizma Base. UDP is measured as UTP after its conversion in an incubation with excess ATP and nucleoside diphosphate kinase. Similarly, UMP is analyzed after it is converted to UDP by nucleoside monophosphate kinase. The uracil nucleotide content of germinated wheat embryos had been determined with this method.
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