Abstract
A rapid method of sequentially phosphorylating picomole quantities of [ 3H]-araC to [ 3H]araCTP is described (ara = 1-β- d-arabinofuranosyl). The procedure utilizes a system of phosphorylating enzymes isolated from rat spleen and requires a single incubation step. The [ 3H]araCTP product is isolated by ion-exchange chromatography and analyzed by PEI-cellulose thin-layer chromatography. At low concentrations of [ 3H]araC as much as 80% can be phosphorylated to the triphosphate, and the produet may be obtained in radiochemical purity greater than 97%.
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