We have examined the use of pulsed-field gel electrophoresis (PFGE) to measure DNA double-strand breaks induced in CHO cells by ionizing radiation. The PFGE assay provides a simple method for the measurement of DNA double-strand breaks for doses as low as 3-4 Gy ionizing radiation, and appears applicable for the measurement of damage produced by any agent producing double-strand breaks. The conditions of transverse alternating field electrophoresis determined both the sensitivity of the assay and the ability to resolve DNA fragments with different sizes. For example, with 0.8% agarose and a 1-min pulse time at 250 V for 18 h of electrophoresis, 0.39% of the DNA per gray migrated into the gel, and only molecules < 1500 kb could be resolved. With 0.56% agarose and a 60-min pulse time at 40 V for 6 days of electrophoresis, 0.55-0.90% of the DNA per gray migrated into the gel, and molecules between 1500 and 7000 kb could be resolved.
Measurement of Radiation-Induced DNA Double-Strand Breaks by Pulsed-Field Gel Electrophoresis
- Views Icon Views
- Share Icon Share
- Search Site
D. D. Ager, W. C. Dewey, K. Gardiner, W. Harvey, R. T. Johnson, C. A. Waldren; Measurement of Radiation-Induced DNA Double-Strand Breaks by Pulsed-Field Gel Electrophoresis. Radiat Res 1 May 1990; 122 (2): 181–187. doi: https://doi.org/10.2307/3577604
Download citation file: