OxiSelect™ Comet Assay Kit (3-Well Slides)User Manual
|Research Area:||Oxidative Stress|
|Assay Category:||DNA/RNA Damage and Repair Assays|
|Detection Method:||Fluorescence Microscopy|
- Useful screening tool for various types of DNA damage
- Slides are specially treated for adhesion of low-melting agarose
- Easy visualization by epifluorescence microscopy
The comet assay, or single cell gel electrophoresis assay (SCGE), is a common technique for measurement of DNA damage in individual cells. Under an electrophoretic field, damaged cellular DNA (containing fragments and strand breaks) is separated from intact DNA, yielding a classic “comet tail" shape under the microscope. Extent of DNA damage is usually visually estimated by comet tail measurement; however, image analysis software is also available for measuring various parameters. The OxiSelect Comet Assay is a fast and sensitive kit for the measurement of cellular DNA damage.
The OxiSelect Comet Assay is a single cell gel electrophoresis assay (SCGE) for simple evaluation of cellular DNA damage. First, individual cells are mixed with molten agarose before application to the OxiSelect Comet Slide. These embedded cells are then treated with a lysis buffer and alkaline solution, which relaxes and denatures the DNA. Finally, the samples are electrophoresed in a horizontal chamber to separate intact DNA from damaged fragments. Following electrophoresis, the samples are dried, stained with a DNA dye, and visualized by epifluorescence microscopy. Under these conditions, the damaged DNA (containing cleavage and strand breaks) will migrate further than intact DNA and produce a “comet tail" shape.
Each kit provides sufficient reagents to perform up to 75 assays.
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