A simple technique for quantitation of low levels of DNA damage in individual cells

X-rays and H202, at relatively low doses, we have demonstrated the detection of DNA damage in individual human lymphocytes

Narendra P. Singh; Michael T. McCoy; Raymond R. Tice; Edward L. Schneider

2006

Scholarcy highlights

  • Human lymphocytes were either exposed to X-irradiation or treated with HzOz at 4°C and the extent of DNA migration was measured using a singlecell microgel electrophoresis technique under alkaline conditions
  • Rydberg and Johanson were the first to directly quantitate DNA damage in individual cells by lysing cells embedded in agarose on slides under mild alkali conditions to allow the partial unwinding of DNA
  • Under the electrophoretic conditions used, no migration of DNA occurred among the majority of the control cells and an approximately linear increase in the length of DNA migration was observed for doses between 25 and 100 rads
  • By 200 rads, the length of migration appeared to plateau, while the extent of DNA damage in cells exposed to greater doses was too great to permit an accurate measurement of the migration pattern
  • In contrast to the relatively homogeneous DNA migration patterns observed for lymphocytes exposed to X-rays, extensive differences in the length of DNA migration, and in the extent of DNA damage, were observed among cells exposed to H202
  • X-rays and H202, at relatively low doses, we have demonstrated the detection of DNA damage in individual human lymphocytes
  • Since the length of migration depends upon the percentage of agarose in the gel and upon the duration of electrophoresis, it should be possible by using either higher-percentage agarose gels or shorter electrophoretic times to quantitate greater amounts of DNA damage in single cells

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