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Additional info for Chromosome Aberrations (Reprint of Cytogenetic and Genome Research 2004)
1A. The plot in Fig. 1A describes the mass of randomly cleaved DNA as a function of Mr. When the X-axis is changed to log10 of Mr, then the plot accurately describes the DNA mass as it is size-fractionated in the log-linear mobility range of agarose gel electrophoresis. , 1999). Using gel electrophoresis of radiolabeled DNA, one can slice the gel and, after scintillation counting, obtain a linear response to the 3[H]DNA content of each gel slice. Cleavage of tritiated V79 hamster cell DNA with EcoRI or HhaI produced mobility distributions that fit a Kuhn distribution (Holmquist, 1988).
After irradiation, the isotonic salt solution was replaced with fresh medium. Cells were periodically harvested over a two-day repair period. Mus spretus × M. castaneus F1 neonatal skin fibroblasts were obtained by collagenase digestion and cultured in DMEM medium + 10 % fetal calf serum. Using a cytogenetic analysis of metaphase spreads, cell population showed 10 % aneuploid cells after 5 days in culture and 100 % aneuploid cells after 14 days in culture. After 20 days, the “post-crisis” fibroblasts were replated.
P values in breaks/kb from Fig. 4 (filled circles) and Fig. 5 (filled squares in the 8 h data) were used. 018 breaks/kb, was subtracted from each P value from irradiated cells to determine the additional breaks attributable to UV irradiation followed by T4-pdg digestion. Global repair follows the first-order kinetics shown wherein CPDs in R-bands disappear with a half-life of about 7 h. Fig. 7. A CPD repair panel from 10 J/m2 UVC-irradiated murine fibroblasts was prepared as in Fig. 2. A murine B2 repeat was used to probe R-band DNA.