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Molecular Approaches For Detecting DNA Damage
P. L. Olive
Published 1998 · Chemistry
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In 1966, McGrath and Williams (51) introduced alkaline sucrose gradient sedimentation as a method to measure DNA single-strand breaks (SSBs) in irradiated cells. This technique was regarded as an important innovation, and was soon in widespread use for examining DNA damage caused by a variety of agents, as well as for measurement of the kinetics of strand break rejoining as an indicator of DNA “repair.” Alkaline sucrose gradient sedimentation remains somewhat unique in that the physical principle behind the measurement of DNA damage is well understood. Unfortunately, the original method proved insensitive to detection of strand breaks produced by “biologically relevant” doses in mammalian cells, largely because conditions required for “xideal” sedimentation required that DNA already contain a significant number of strand breaks. Ability to analyze only a few samples at a time and the long time required for the assay were also viewed as serious limitations.
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