Cell-cycle variation in DNA migration in pulsed-field gel electrophoresis

Abstract

Pulsed-field electrophoresis is being used extensively in the gene mapping studies and in the analysis of DNA strand breakage by ionizing radiation. We have evaluated the relationship between the fraction of S phase DNA in a cell population and its ability to modify the migration of DNA in pulsed-field gel electrophoresis. We have shown that increasing the proportion of S phase DNA reduced the effective rate of migration of MGH-U1 cellular DNA. This effect was observed after treatment with ionizing radiation or the restriction enzyme Not I. However, when radiationinduced damage was studied using intact cells, only the DNA with 70% S phase showed apparent differences in damage induction. These studies therefore provide data to indicate the percentage of S phase cells at which overall DNA migration might be affected significantly.