Twelve-gel slide format optimised for comet assay and fluorescent in situ hybridisation

Functional, Genetic, and Epigenetic Aspects of Base and Nucleotide Excision Repair in Colorectal Carcinomas

“…In vitro repair assays, adopted from Langie and colleagues (20,21), were implemented using a 12-gel slide format (22). Briefly, protein extracts were incubated with 2 types of substrate DNA, containing artificially induced lesions known to be repaired specifically by BER or NER pathway.…”

Comet Assay on Toxicogenetics; Several Studies in Recent Years on Several Genotoxicological Agents

“…In another study, a high-throughput Comet Chip screening assay based on a microfabricated 96-well design with automated processing was used to evaluate DNA damage caused by ZnO, Ag, Fe 2 O 3 , CeO 2 , and SiO 2 NPs in human lymphoblastoid (TK6) cells in suspension, and adherent Chinese hamster ovary (H9T3) cells [25]. High throughput comet assay approaches, whether 12 mini-gels on a slide, 48 or 96 mini-gels on a GelBond film [6,7,24], or microfabricated 96-well CometChips [25], all show robustness, improved efficiency and reduced processing time. In addition, they reduce the risk of user bias in comparison with the standard comet assay (Table 1 and Figure 2).…”

“…It allows investigation of DNA damage in virtually all cell types without the necessity of cell cultures [2]. It is widely used to detect DNA damage [5] as an indicator of exposure to genotoxicogical agents [2,6,7]. The Comet assay is a used method in human, environmental, and ecogenotoxicological studies [2] and it is performed to detect genotoxicity effect of biocides, chemicals prodcuts, agrochemicals, pharmaceuticals and food additives in genotoxicity assaying [8].…”

Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

“…Cells were resuspended in PBS at 0.25x10 6 cells/ml. 20 μl of cell suspension was mixed with 94 μl of 1% low-melting point agarose, and 5 µl drops were placed onto agarose-precoated slides following a format of 12 minigels/slide [31]. Cells were lysed in 2.5 M NaCl, 0.1 M EDTA, 0.01 M Tris and 1% Triton X-100 (pH 10) at 4°C for at least 1 h. After that, slides were immersed in electrophoresis solution (0.3 M NaOH and 1 mM EDTA) for 20 min.…”

“…Using a fluorescence microscope we visually classified 100 comets (50 on each gel) into 5 categories, 0-4, representing increasing relative tail intensities. Summing the scores (0-4) of 100 comets therefore gives an overall score of between 0 and 400 arbitrary units [31].…”