Single-strand breaks are a major form of DNA damage caused by ionizing radiation, and measurement of strand breaks has long been used as an index of overall cellular DNA damage. Most assays for DNA single-strand breaks in cells rely on measuring fractionated DNA samples following alkali denaturation. Quantification is usually achieved by prelabeling cells with radioactive DNA precursors; however, this is not possible in the situation of nondividing cells or freshly isolated tissue. It has previously been demonstrated that the alkali unwinding assay of DNA strand breaks can be quantified by blotting the recovered DNA on nylon membranes and hybridizing with radiolabeled sequence-specific probes. We report here improvements to the technique, which include hot alkali denaturation of DNA samples prior to blotting and the use of carrier DNA that is noncomplementary to the radiolabeled probe. Our method allows both single- and double-stranded DNA to be quantified with the same efficiency, thereby improving the sensitivity and reproducibility of the assay, and allows calibration for determination of absolute levels of DNA strand breaks in cells. We also used this method to assay radiation-induced DNA strand breaks in freshly isolated human leukocytes and found them to have a strand break induction rate of 1815 strand breaks/cell/Gy.
Skip Nav Destination
Research Article| November 01 1992
Radiation-Induced DNA Single-Strand Breaks in Freshly Isolated Human Leukocytes
John M. Buatti ;
Luis R. Rivero ;
Radiat Res (1992) 132 (2): 200–206.
- Views Icon Views
- Share Icon Share
- Search Site
John M. Buatti, Luis R. Rivero, Timothy J. Jorgensen; Radiation-Induced DNA Single-Strand Breaks in Freshly Isolated Human Leukocytes. Radiat Res 1 November 1992; 132 (2): 200–206. doi: https://doi.org/10.2307/3578527
Download citation file:
Citing articles via
Commonalities Between COVID-19 and Radiation Injury
Carmen I. Rios, David R. Cassatt, Brynn A. Hollingsworth, Merriline M. Satyamitra, Yeabsera S. Tadesse, Lanyn P. Taliaferro, Thomas A. Winters, Andrea L. DiCarlo
Low-Dose Radiation Therapy (LDRT) for COVID-19: Benefits or Risks?
Pataje G. Prasanna, Gayle E. Woloschak, Andrea L. DiCarlo, Jeffrey C. Buchsbaum, Dörthe Schaue, Arnab Chakravarti, Francis A. Cucinotta, Silvia C. Formenti, Chandan Guha, Dale J. Hu, Mohammad K. Khan, David G. Kirsch, Sunil Krishnan, Wolfgang W. Leitner, Brian Marples, William McBride, Minesh P. Mehta, Shahin Rafii, Elad Sharon, Julie M. Sullivan, Ralph R. Weichselbaum, Mansoor M. Ahmed, Bhadrasain Vikram, C. Norman Coleman, Kathryn D. Held
Germicidal Efficacy and Mammalian Skin Safety of 222-nm UV Light
Manuela Buonanno, Brian Ponnaiya, David Welch, Milda Stanislauskas, Gerhard Randers-Pehrson, Lubomir Smilenov, Franklin D. Lowy, David M. Owens, David J. Brenner
Photon GRID Radiation Therapy: A Physics and Dosimetry White Paper from the Radiosurgery Society (RSS) GRID/LATTICE, Microbeam and FLASH Radiotherapy Working Group
Hualin Zhang, Xiaodong Wu, Xin Zhang, Sha X. Chang, Ali Megooni, Eric D. Donnelly, Mansoor M. Ahmed, Robert J. Griffin, James S. Welsh, Charles B. Simone, II, Nina A. Mayr