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- W90138503 abstract "A method is described in detail for measuring repair replication, which has proven useful in studies with bacteria, as well as many eukaryotes, including protozoans and mammalian cells. The method has also been utilized to study repair replication in permeabilized systems to which the nucleoside triphosphates and macromolecules may be added. Cellular DNA is prelabeled by growth in a medium containing /sup 14/C- or /sup 32/P-labeled DNA precursors. The cells are treated with the test agent and incubated in a medium containing the density label 5-bromouracil or 5-bromodeoxyuridine, which is /sup 3/H-labeled to radioactively label the newly synthesized DNA. After further incubation, the cells are harvested, lysed, and treated with protease. The digest is made to a density of 1.72 g ml /sup -1/ with CsCl. The solution is centrifuged to equilibrium, and fractions are collected and assayed for radioactivity. The fractions containing unreplicated parental density DNA are rebanded in a second CsCl equilibrium density gradient to further resolve repair-synthesized DNA from semi-conservatively replicated DNA. The ratio of /sup 3/H to /sup 32/P of /sup 14/C in the DNA of parental density in the second gradient provides a measure of repair." @default.
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- W90138503 date "1981-01-01" @default.
- W90138503 modified "2023-09-27" @default.
- W90138503 title "Measurement of repair replication by equilibrium sedimentation" @default.
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