Towards a more reliable comet assay: Optimising agarose concentration, unwinding time and electrophoresis conditions
Towards a more reliable comet assay: Optimising agarose concentration, unwinding time and electrophoresis conditions
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DOI：
10.1016/j.mrgentox.2011.05.010
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发表时间：
2011-09-18
影响因子：
1.9
通讯作者：
Collins, Andrew R.
中科院分区：
文献类型：
--
作者：
Azqueta, Amaya;Gutzkow, Kristine B.;Collins, Andrew R.
彗星试验现在是测量细胞中大多数类型的DNA损伤的首选方法。然而，由于缺乏标准化的方案，实验室间的比对价值有限。这篇论文的目的是论证彗星分析变量的微小变化如何显著影响结果。我们用两种细胞类型，即人外周血淋巴细胞和淋巴母细胞系TK-6，研究了不同琼脂浓度、碱性解离时间、电泳时间、电压和电流的影响。所有这些变量都对分析性能有显著影响，因此对DNA损伤的确定也有显著影响。在这里，我们确定了特别重要的因素。(C)2011爱思唯尔B.V.保留所有权利。
The comet assay is now the method of choice for measuring most kinds of DNA damage in cells. However, due to the lack of a standardised protocol inter-laboratory comparisons are of limited value. The aim of this paper is to demonstrate how small changes in comet-assay variables may significantly affect the results. We examined the effect of varying agarose concentrations, alkaline unwinding time, electrophoresis time, voltage and current, by use of two cell types, viz, human peripheral blood lymphocytes and the lymphoblastoid cell line TK-6. All these variables have marked effects on assay performance and, therefore, on the determination of DNA damage. Here we identify factors of particular importance. (C) 2011 Elsevier B.V. All rights reserved.