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A rapid quantitative determination of deoxyribonucleoside triphosphates based on the enzymatic synthesis of DNA

1969; Elsevier BV; Volume: 174; Issue: 2 Linguagem: Inglês

10.1016/0005-2787(69)90288-3

1879-3002

Alan W. Solter, Robert E. Handschumacher,

RNA and protein synthesis mechanisms

An assay has been developed for deoxyribonucleoside triphosphates; it is based on the observation that all four nucleotides are necessary for the continued synthesis of DNA. A linear standard curve is obtained with one of the nucleotides as a limiting factor and measurement of the amount of incorporation into the polymer of a radioactive deoxyribonucleoside triphosphate present in excess. The method has been utilized to determine the concentration of dTTP in HClO4 extracts of mouse leukemia cells and Streptococcus faecalis. Other materials present in the extracts did not influence the assay. The method offers the advantages of simplicity, speed, and sensitivity.