Department of Chemicals and Radiation, National Institute of Public Health, PO Box 4404 Nydalen, Oslo N-0403, Norway.
Mutagenesis. 2013 May;28(3):333-40. doi: 10.1093/mutage/get012. Epub 2013 Mar 5.
The single-cell gel electrophoresis--the comet assay--has proved to be a sensitive and relatively simple method that is much used in research for the analysis of specific types of DNA damage, and its use in genotoxicity testing is increasing. The efficiency of the comet assay, in terms of number of samples processed per experiment, has been rather poor, and both research and toxicological testing should profit from an increased throughput. We have designed and validated a format involving 96 agarose minigels supported by a hydrophilic polyester film. Using simple technology, hundreds of samples may be processed in one experiment by one person, with less time needed for processing, less use of chemicals and requiring fewer cells per sample. Controlled electrophoresis, including circulation of the electrophoresis solution, improves the homogeneity between replicate samples in the 96-minigel format. The high-throughput method described in this paper should greatly increase the overall capacity, versatility and robustness of the comet assay.
单细胞凝胶电泳 - 彗星试验 - 已被证明是一种灵敏且相对简单的方法，广泛用于分析特定类型的 DNA 损伤，并且其在遗传毒性测试中的应用正在增加。彗星试验在每个实验中处理的样品数量方面的效率相当差，并且研究和毒理学测试都应该从增加的通量中受益。我们设计并验证了一种涉及 96 个琼脂糖微型凝胶的格式，由亲水聚酯膜支撑。使用简单的技术，一个人可以在一个实验中处理数百个样品，处理时间更短，化学品使用更少，每个样品所需的细胞更少。控制电泳，包括电泳溶液的循环，可提高 96 微型凝胶格式中重复样品之间的均匀性。本文所述的高通量方法应大大提高彗星试验的整体容量、多功能性和稳健性。
