OxiSelect™ Comet Assay Kit (3-Well Slides)

Protocol tips

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Seed 1x10^5 cells/ml		The DNA damage is quantified by measuring the displacement between the genetic material of the nucleus ('comet head') and the resulting 'tail'. Tail Moment and Tail DNA% are the two most common parameters to analyze Comet assay results. At least 50 -100 cells should be analyzed per sample. The Tail Moment has been suggested to be an appropriate index of induced DNA damage in considering both the migration of the genetic material as well as the relative amount of DNA in the tail.

Upstream tips
Seed 1x10^5 cells/ml
Downstream tips
The DNA damage is quantified by measuring the displacement between the genetic material of the nucleus ('comet head') and the resulting 'tail'. Tail Moment and Tail DNA% are the two most common parameters to analyze Comet assay results. At least 50 -100 cells should be analyzed per sample. The Tail Moment has been suggested to be an appropriate index of induced DNA damage in considering both the migration of the genetic material as well as the relative amount of DNA in the tail.

Publication protocol

Using the OxiSelectTM 96-Well Comet Assay Kit (Cell Biolabs, San Diego, CA, USA) according to the manufacturer’s instructions, we trypsinized cells treated with 3-MCPD for 24 hr, then diluted with ice-cold PBS. The positive controls here are 100 μM H2O2-treated cells. We mixed 3-MCPD- and H2O2-treated cells with low-melting-point agarose to coat the slides. Next, we treated the slides with lysis buffer and alkaline solution, electrophoresed them under neutral conditions, and stained them with DNA dye. For analysis, we selected two slides per concentration and a total of 50 comets per group (25 cells per slide). CASP image analysis software was applied to the captured images. We measured the olive tail moment (OTM) (Kent et al., 1995), calculated as comet tail moment length multiplied by the relative DNA content of the tail, as an indicator of DNA damage.

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Download the product protocol from Cell Biolabs for OxiSelect™ Comet Assay Kit (3-Well Slides) below.

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