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Single cell gel electrophoresis comet assay kit

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The Single cell gel electrophoresis (comet assay) kit is a laboratory equipment designed to measure DNA damage and repair in individual cells. The kit provides the necessary materials and protocols to perform the comet assay, a widely used technique for evaluating genotoxicity and cellular response to DNA-damaging agents.

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Lab products found in correlation

Dna isolation kit, by Omega Bio-Tek (1 mentions) Diaphot 200, by Nikon (1 mentions)

Close spelling variants of this product

Comet Assay Single Cell Gel Electrophoresis Assay Kit Comet assay kit for single cell gel electrophoresis Single Cell Gel Electrophoresis Assay Kit Comet Assay™ electrophoresis kit Comet Assay Kit

3 protocols using single cell gel electrophoresis comet assay kit

1

Comet Assay for DNA Damage Quantification

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A single cell gel electrophoresis (comet assay) kit was employed to evaluate DNA damage (Trevigen, Inc., Gaithersburg, MD, USA). After the treatment of cells with Cis, the genomic DNA was purified using a DNA isolation kit (Omega Bio-Tek, Inc., Norcross, GA, USA). Subsequently, the amount of DNA damage was quantified based on comet tail lengths (consisting of the nuclear region and tail) which were scored visually as described in previous work.15 (link)
Liu A.S., Yu H.Y., Yang Y.L., Xue F.Y., Chen X., Zhang Y., Zhou Z.Y., Zhang B., Li L., Sun C.Z., Huang P, & Huang J.F. (2019). A Chemotherapy-Driven Increase in Mcl-1 Mediates the Effect of miR-375 on Cisplatin Resistance in Osteosarcoma Cells. OncoTargets and therapy, 12, 11667-11677.
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2

Comet Assay for DNA Damage Assessment in U87 Cells

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To determine the extent of DNA damage, individual U87 cells were treated with a vehicle (0.05% dimethyl sulfoxide [DMSO]) or 10 and 30 μM SFN, respectively, for 24 hours, and the cells were processed for the comet assay under alkaline as well as neutral conditions, as previously described,48 (link) using the single cell gel electrophoresis/comet assay kit (Trevigen). Comets were visualized by epifluorescence microscopy using a fluorescein isothiocyanate filter. The slides were stained with SYBR Green for 5 minutes while protected from light and dried at room temperature before visualization. Fluorescence images were captured using a Nikon Diaphot 200 fluorescence microscope. The comet lengths of the individual cells per group were measured in each treatment group. Two hundred comets were randomly analyzed by measuring the distance of DNA migration in each cell from the nuclear core to the trailing edge of the comet using TriTek CometScore TM Freeware v1.5C software (TriTek Corporation).
Bijangi-Vishehsaraei K., Saadatzadeh M.R., Wang H., Nguyen A., Kamocka M.M., Cai W., Cohen-Gadol A.A., Halum S.L., Sarkaria J.N., Pollok K.E, & Safa A.R. (2017). Sulforaphane suppresses the growth of glioblastoma cells, glioblastoma stem cell–like spheroids, and tumor xenografts through multiple cell signaling pathways. Journal of neurosurgery, 127(6), 1219-1230.
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3

Comet Assay for DNA Damage Assessment

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A single-cell gel electrophoresis (comet assay) kit was employed for evaluating DNA damage (Trevigen, Inc.). Etoposide- or UV-treated or untreated MCF7 cells were resuspended in PBS and combined with molten low melting agarose at a ratio of 1:10 (v/v) and immediately pipetted 50 μl onto CometSlide. After cell lysis, samples were treated with NaOH solution (300 mM NaOH, 1mM EDTA) and applied to electrophoresis. Then the samples were stained with DAPI, and viewed by a fluorescence microscopy [50 (link)]. Images were analysed with an ImageJ software plugin Comet Assay from Microscopy Services Laboratory (https://www.med.unc.edu/microscopy) [57 (link)].
Xia L., Qin K., Wang X.R., Wang X.L., Zhou A.W., Chen G.Q, & Lu Y. (2017). Pyruvate kinase M2 phosphorylates H2AX and promotes genomic instability in human tumor cells. Oncotarget, 8(65), 109120-109134.
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