Contamination by ATP of commercial dATP samples causing erroneous results in studies of DNA replication in isolated HeLa cell nuclei.

Abstract

dATP at high concentrations was capable of replacing ATP required in the synthesis of Okazaki pieces in isolated HeLa cell nuclei. In addition, the levels of synthesis of high molecular weight DNA were observed to vary depending on the lot of dATP used. Analysis by HPLC revealed that dATP samples of a particular source contained ATP in the range of 0.25-0.43 mol%. With ATP-free dATP, almost no synthesis of high molecular weight DNA was observed, while with impure dATP, a small but significant amount of high molecular weight DNA was synthesized. While this observation confirmed our previous finding that dATP can replace ATP in the synthesis of Okazaki pieces but not in the joining of the pieces, it is also a warning to users of commercial dATP in biochemical and biological studies.