Oxiselect Comet Assay Kit

Background: DNA damage, due to environmental factors and normal metabolic processes inside the cell, occurs at a rate of 1,000 to 1,000,000 molecular lesions per cell per day. While this counts for only a small part of the human genome's approximately 6 billion bases (3 billion base pairs), unrepaired lesions to critical genes can impede a cell's ability to carry out its function and appreciably increase the likelihood of cancer. 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. Each kit provides sufficient reagents to perform up to 75 assays.

Principle of the Assay: 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 (see Figure 1). Each kit provides sufficient reagents to perform up to 75 assays.

1. Ostling, O., and Johanson, K. J. (1984). Biochem. Biophys. Res. Commun. 123, 291-298.
2. Singh, N. P., McCoy, M. T., Tice, R. R., and Schneider, E. L. (1988). Exp. Cell. Res. 175, 184-191.
3. Olive, P. L., Banath, J. P., and Durand, R. E. (1990). Radiat. Res. 122, 86-94.
4. De Boeck, M., Touil, N., De Visscher, G., Vande, P. A., and Kirsch-Volders, M. (2000). Mutat. Res. 469, 181-197
1. Wang, L. et al. (2014). Osmotic Stress-induced Phosphorylation of H2AX by Polo-like Kinase 3 Affects Cell Cycle Progression in Human Corneal Epithelial Cells. J Biol Chem. 289:29827-29835.
2. Savage, K.I. et al. (2014). BRCA1 Deficiency Exacerbates Estrogen-Induced DNA Damage and Genomic Instability. Cancer Res. 74:2773-2784.
3. Robin, T.P. et al. (2012). EWS/FLI1 Regulates EYA3 in Ewing Sarcoma via Modulation of miRNA-708, Resulting in Increased Cell Survival and Chemoresistance. Mol. Cancer Res. 10:1098-1108.
4. Choi, S.K. et al. (2012).Poly(ADP-Ribose) Polymerase 1 Inhibition Improves Coronary Arteriole Function in Type 2 Diabetes Mellitus. Hypertension. 59:1060-1068.

Small volumes of OxiSelect Comet assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS168415 is an assay kit targeting OxiSelect Comet. Also known as OxiSelect Comet Assay Kit (3-Well Slides).