The rapid, simple and improved preparation of high specific activity alpha-[32P]dATP and alpha-[32P]ATP.
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 enzymic conversion to the triphosphate with myokinase (EC 2.7.4.3) and pyruvate kinase (EC 2.7.1.40) 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.
D000255:Adenosine Triphosphate; D000263:Adenylate Kinase; D003838:Deoxyadenine Nucleotides; D007553:Isotope Labeling / Q000379:methods; D010761:Phosphorus Radioisotopes; D011770:Pyruvate Kinase
D003838:Deoxyadenine Nucleotides; D010761:Phosphorus Radioisotopes; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; D000263:Adenylate Kinase