Hunting D, Henderson J F
Can J Biochem. 1981 Sep;59(9):723-7. doi: 10.1139/o81-100.
There are more than twenty appreciably different deoxyribonucleoside triphosphate assays using DNA polymerase in the literature. Therefore, each aspect of this method has been critically evaluated, including the purity of the substrates and of DNA polymerase, the reaction conditions, product isolation, and the effect of cell extracts on the assay. The final method uses a phosphatase-free DNA polymerase preparation, 2'-dAMP to inhibit the 3' leads to 5' exonuclease of DNA polymerase I, and includes a correction of both the background incorporation and the sample incorporation for dilution of the specific activity of the radioactive precursor by the sample. The sensitivity, range, accuracy, and reproducibility are reported as well as values for the deoxyribonucleoside triphosphate content of Chinese hamster ovary K-1 cells.
文献中有二十多种使用DNA聚合酶的明显不同的脱氧核糖核苷三磷酸测定方法。因此，对该方法的各个方面都进行了严格评估，包括底物和DNA聚合酶的纯度、反应条件、产物分离以及细胞提取物对测定的影响。最终方法使用不含磷酸酶的DNA聚合酶制剂、2'-dAMP抑制DNA聚合酶I的3'→5'外切核酸酶，并对背景掺入和样品掺入进行校正，以校正样品对放射性前体比活性的稀释。报告了该方法的灵敏度、范围、准确性和重现性以及中国仓鼠卵巢K-1细胞的脱氧核糖核苷三磷酸含量值。
