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Nucleic Acids Research, 1977, Vol. 4, No. 12 4347-4356
© 1977

Articles

The rapid, simple and improved preparation of high specific activity {alpha}-[32P]dATP and {alpha}-[32p]ATP

Robert H. Symons

Department of Biochemistry, University of Adelaide Adelaide, South Australia, 5001

Received October 18, 1977. An improved method is described for the rapid and simple preparation of {alpha}-[32P]dATP and {alpha}-32P]ATP from 32Pi in good yields and with specific activities from 20 – 150 Ci/mmol. The two-step procedure involves the chemical synthesis of the mononucleotide followed by its enzyinic conversion to the triphosphate with myokinase (EC 2.7.4.3 [EC] ) and pyruvate kinase (EC 2.7.1.40 [EC] ) in the presence of trace amounts of dATP or ATP to prime the reaction. The two steps are carried out in the same reaction flask and the only purification step required is a step-wise elution from a column of DEAE-cellulose.


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