Detection of DNA strand breaks in barley roots after .GAMMA.-ray irradiation using the alkaline unwinding assay.

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  • SHIKAZONO NAOYA
    Laboratory of Radiation Genetics and Chemical Mutagenesis, Faculty of Agriculture, The University of Tokyo
  • WATANABE HIROSHI
    Biotechnology Laboratory, Department of Radiation Research for Environment and Resources, Japan Atomic Energy Research Institute
  • TANAKA ATSUSHI
    Advanced Science Research Center, Japan Atomic Energy Research Insitute
  • TANO SHIGEMITSU
    Advanced Science Research Center, Japan Atomic Energy Research Insitute
  • HIRAI ATSUSHI
    Laboratory of Radiation Genetics and Chemical Mutagenesis, Faculty of Agriculture, The University of Tokyo

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Description

The alkaline unwinding assay was applied to measure strand breaks in DNA of intact barley roots after γ-ray irradiation without extracting DNA from the tissue. The most suitable condition for unwinding was determined to be 0.03 M NaOH containing 0.5 M NaCl for alkaline solution composition, and 15 min at 20°C for unwinding time. Under this condition, DNA strand breaks were shown to be linearly increased with increasing dose, and further be rejoined during the post-irradiation incubation for 6 hr.

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