Comet Assay Kits, 96-Well

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Seed 1 × 10^5 cells /ml

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Seed 1 × 10^5 cells /ml

Publication protocol

Single cell gel electrophoresis (SCGE) was carried out by Trevigen’s comet assay kit with slight modification. Briefly, HepG2 cell suspension (1 × 10^5/ml) and molten LM agarose (at 37°C) were prepared in 1:10 (v/v) ratio. From there, instantly 50 μl was poured onto the comet slide pre-coated with 1% normal melting point agarose. Then, the slides were kept at 4°C in the dark for 10 min under low humidity and dust-free environment for their better agarose adhesion on the slides. The slides were placed in an ice-cold lysing solution for 60 min at 4°C. Furthermore, the excess buffer of slides was drained and deeply immersed in freshly prepared alkaline unwinding solution (pH>13: 200 mM NaOH, 1 mM EDTA in 50 ml of dH2O) for 1 h at 4°C in the dark. Subsequently, the slides were placed in an electrophoresis chamber filled with alkaline buffer (200 mM NaOH, 1 mM EDTA in 1 l of dH2O) and it runs with 21 V for 30 min., Excess electrophoresis solution from the slides was drained gently and then a couple time immersed in dH2O and 70% ethanol for each 5 min. Eventually, slides were dried for 10–15 minutes at room temperature and stored with desiccant prior to scoring slides. Silver staining was performed to visualize the comets through transmission light microscope. Randomly, 50 comets per slide were visualized and analyzed by scoring of various parameters such as head, tail and tail moment under light microscope with comet assay software.

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