A simple method for the preparation of [beta-32P]purine nucleoside triphosphase

Nucleic Acids Res. 1977 Oct;4(10):3341-55. doi: 10.1093/nar/4.10.3341.

Abstract

A rapid, simple and inexpensive procedure is described for the preparation of purine ribo-and deoxyribonucleoside triphosphates specifically and highly radiolabeled with [32P]phosphate in the beta position. The method involves two successive enzymatic reactions: conversion of donor [gamma-32P]ATP in the presence of an excess of acceptor 5'-mononucleotide to the 5'-diphosphates by myokinase or guanosine 5'-monophosphate kinase followed by phosphorylation with pyruvate kinase to yield 5'-triphosphates.

MeSH terms

  • Adenylate Kinase
  • DNA-Directed RNA Polymerases / metabolism
  • Isotope Labeling / methods
  • Phosphoenolpyruvate Carboxykinase (GTP)
  • Phosphorus Radioisotopes
  • Purine Nucleotides* / biosynthesis
  • RNA, Messenger / biosynthesis
  • Reoviridae / enzymology
  • Vaccinia virus / enzymology

Substances

  • Phosphorus Radioisotopes
  • Purine Nucleotides
  • RNA, Messenger
  • Adenylate Kinase
  • DNA-Directed RNA Polymerases
  • Phosphoenolpyruvate Carboxykinase (GTP)