Synthesis of (alpha-32P) ribo- and deoxyribonucleoside 5'-triphosphates.

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Publisher Summary For the routine preparation of the common [α- 32 P]ribo- and deoxyribonucleoside 5'-triphosphates with high specific activity, the most practical route is the chemical synthesis of the monophosphate from [ 32 P]orthophosphoric acid and the appropriate nucleoside followed by its enzymatic conversion to the triphosphate. This chapter describes the method for the synthesis of [α-32P] ribo- and deoxyribonucleoside 5'-triphosphates. This method allowed the preparation of [a- 32 P]ribo- and deoxyribonucleoside 5'-triphosphates with a specific activity of 1-20 mCi/μmole, higher specific activities are quite feasible. A total working time of about 3 hours spread over three days is required for the preparation of each labeled triphosphate. It is considered desirable to purify reaction mixtures after the chemical synthesis of the monophosphate as well as after the enzymatic synthesis of the triphosphate in order to ensure a high purity of the final product. However, purificatio

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