Computer simulation of pulsed field gel runs allows the quantitation of radiation-induced double-strand breaks in yeast.
A procedure for the quantification of double-strand breaks in yeast is presented that utilizes pulsed field gel electrophoresis (PFGE) and a comparison of the observed DNA mass distribution in the gel lanes with calculated distributions. Calculation of profiles is performed as follows. If double-strand breaks are produced by sparsely ionizing radiation, one can assume that they are distributed randomly in the genome, and the resulting DNA mass distribution in molecular length can be predicted by means of a random breakage model. The input data for the computation of molecular length profiles are the breakage frequency per unit length, alpha, as adjustable parameter, and the molecular lengths of the intact chromosomes. The obtained DNA mass distributions in molecular length must then be transformed into distributions of DNA mass in migration distance. This requires a calibration of molecular length vs. migration distance that is specific for the gel lane in question. The computed profiles are then folded with a Lorentz distribution with adjusted spread parameter gamma to account for band broadening. The DNA profiles are calculated for different breakage frequencies alpha and for different values of gamma, and the parameters resulting in the best fit of the calculated to the observed profile are determined.
GSF-Forschungszentrum, Institut für Strahlenbiologie, Neuherber, Germany.
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Kraxenberger
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Friedl
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AM
Kellerer
D003198:Computer Simulation; D004249:DNA Damage; D004271:DNA, Fungal / Q000032:analysis* / Q000528:radiation effects; D016521:Electrophoresis, Gel, Pulsed-Field; D005720:Gamma Rays; D007091:Image Processing, Computer-Assisted; D015233:Models, Statistical; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004271:DNA, Fungal