MEASUREMENT OF RADIATION-INDUCED DNA DOUBLE-STRAND BREAKS BY PULSED-FIELD GEL-ELECTROPHORESIS

被引：112
作者：
AGER, DD
DEWEY, WC
GARDINER, K
HARVEY, W
JOHNSON, RT
WALDREN, CA
机构：
[1] UNIV CAMBRIDGE,DEPT ZOOL,CANC RES CAMPAIGN,MAMMALIAN CELL DNA REPAIR RES GRP,CAMBRIDGE CB2 3EJ,ENGLAND
[2] ELEANOR ROOSEVELT INST CANC RES,DENVER,CO 80206
来源：
RADIATION RESEARCH | 1990年 / 122卷 / 02期
关键词：
D O I：
10.2307/3577604
中图分类号：
Q [生物科学];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
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.
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页码：181 / 187
页数：7
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