Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis
Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis
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DOI：
10.1016/j.mrfmmm.2003.08.005
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发表时间：
2003-10-29
影响因子：
2.3
通讯作者：
Sutherland, JC
中科院分区：
文献类型：
--
作者：
Sutherland, BM;Georgakilas, AG;Sutherland, JC
评估DNA损伤的诱导、修复和这种损伤的后果需要通过独立于对这种损伤的生物反应的方法来测量特定的DNA损伤。影响一条DNA链的损伤（改变的碱基、脱碱基位点、单链断裂（SSB））以及影响两条链的损伤（成簇损伤、双链断裂）可以通过使用凝胶电泳、凝胶成像和数均长度分析直接测量DNA来定量。低至每千兆碱基对（10（9）bp）几个位点的损伤频率可以通过这种方法在约50 ng的非放射性DNA中定量，并且单分子方法可以允许在来自单细胞的DNA中进行这样的测量。本文介绍了这种方法的理论基础，生化要求和实践方面，并展示了他们的应用程序中的识别和定量的复杂的集群损伤的例子。(C)2003 Elsevier B.V.保留所有权利。
Assessing DNA damage induction, repair and consequences of such damages requires measurement of specific DNA lesions by methods that are independent of biological responses to such lesions. Lesions affecting one DNA strand (altered bases, abasic sites, single strand breaks (SSB)) as well as damages affecting both strands (clustered damages, double strand breaks) can be quantified by direct measurement of DNA using gel electrophoresis, gel imaging and number average length analysis. Damage frequencies as low as a few sites per gigabase pair (10(9) bp) can be quantified by this approach in about 50 ng of non-radioactive DNA, and single molecule methods may allow such measurements in DNA from single cells. This review presents the theoretical basis, biochemical requirements and practical aspects of this approach, and shows examples of their applications in identification and quantitation of complex clustered damages. (C) 2003 Elsevier B.V. All rights reserved.