用于检测和测量DNA单链断裂的邻羟基胺偶联碱性凝胶电泳测定法
O-hydroxylamine-coupled alkaline gel electrophoresis assay for the detection and measurement of DNA single-strand breaks.
作者信息
Luke April M, Nakamura Jun
机构信息
Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC, USA.
出版信息
Methods Mol Biol. 2012;920:307-13. doi: 10.1007/978-1-61779-998-3_21.
The ability to detect and measure DNA single-strand breaks has been the aim of numerous assays developed to assess genotoxicity. These methods often rely on alkaline conditions to denature DNA. However, alkaline treatment of DNA also introduces artifactual SSBs through the cleavage of alkali-labile sites resulting in confounded data. Here, we describe a modified alkaline gel electrophoresis assay coupled with a neutral O-hydroxylamine to obtain the measurement of true SSB formation.
检测和测量DNA单链断裂的能力一直是为评估遗传毒性而开发的众多检测方法的目标。这些方法通常依赖碱性条件使DNA变性。然而，对DNA进行碱性处理也会通过切割碱不稳定位点引入人为的单链断裂，从而导致数据混淆。在此，我们描述了一种改良的碱性凝胶电泳检测方法，结合中性O-羟胺来测量真正的单链断裂形成情况。
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