Department of Nutrition, University of Oslo, Blindern, Oslo, Norway.
Toxicol Lett. 2010 May 19;195(1):31-4. doi: 10.1016/j.toxlet.2010.02.017. Epub 2010 Feb 25.
The comet assay is widely used to measure DNA damage and repair in basic research, genotoxicity testing and human biomonitoring. The conventional format has 1 or 2 gels on a microscope slide, 1 sample per slide. To increase throughput, we have designed and tested a system with 12 smaller gels on one slide, allowing incubation of individual gels with different reagents or enzymes. Thus several times more samples can be analysed with one electrophoresis run, and fewer cells and smaller volumes of test solutions are required. Applications of the modified method include treatment with genotoxic agents at different concentrations; simultaneous analysis of different lesions using a range of enzymes; analysis of cell extracts for DNA repair activity; and fluorescent in situ hybridisation (FISH) to comet DNA with specific labelled probes.
彗星实验广泛用于基础研究、遗传毒性测试和人体生物监测中,以测量 DNA 损伤和修复。传统的格式是在显微镜载玻片上有 1 或 2 个凝胶,每个载玻片上有 1 个样本。为了提高通量,我们设计并测试了一种带有 12 个较小凝胶的系统,允许在每个凝胶上用不同的试剂或酶进行孵育。因此,在一次电泳运行中可以分析更多的样本,所需的细胞和测试溶液的体积更小。改良方法的应用包括在不同浓度下用遗传毒性剂处理;使用一系列酶同时分析不同的损伤;分析细胞提取物中的 DNA 修复活性;以及用特定标记探针进行彗星 DNA 的荧光原位杂交 (FISH)。