The rapid, simple and improved preparation of high specific activity α-[32P]dATP and α-[32p]ATP | Zendy

Robert H. Symons | Zendy

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

Author(s) -

Robert H. Symons

Publication year - 1977

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/4.12.4347

Subject(s) - elution , adenylate kinase , biology , chromatography , adenosine triphosphate , specific activity , pyruvate kinase , cellulose , biochemistry , enzyme , chemistry , glycolysis

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.

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