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A rapid enzymatic procedure for production of 1-β-d-[3H]arabinofuranosyl cytosine 5′-triphosphate

✍ Scribed by Carol G. Mintz; Cornelia Agent; N.Burr Furlong

Publisher
Elsevier Science
Year
1976
Tongue
English
Weight
432 KB
Volume
75
Category
Article
ISSN
0003-2697

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✦ Synopsis

A rapid method of sequentially phosphorylating picomole quantities of PHI-araC to [3H]araCTP is described (ara = I-P-D-arabinofuranosyb.

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 product may be obtained in radiochemical purity greater than 97%.

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