Assessment of DNA damage and DSB formation in cells was performed using the CometAssay single cell gel electrophoresis assay (Trevigen). Cells were harvested and resuspended in low-melting point agarose, plated onto provided glass slides, and subjected to electrophoresis in neutral electrophoresis buffer (100 mM Tris, 300 mM Na Acetate, pH 9.0). Slides were processed according to manufacturer’s instructions. DNA tails were visualized after SYBR Gold staining using a Nikon Eclipse 80i fluorescence microscope and quantified using ImageJ software with the OpenComet plugin (URL http://www.opencomet.org ).
Cometassay single cell gel electrophoresis assay
Manufactured by Bio-Techne
The CometAssay single cell gel electrophoresis assay is a laboratory technique used to detect and quantify DNA damage in individual cells. The assay utilizes gel electrophoresis to separate fragments of damaged DNA, which appear as a 'comet-like' shape under a microscope. This method provides a sensitive and reliable way to measure DNA damage and repair at the single-cell level.
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Lab products found in correlation
2 protocols using cometassay single cell gel electrophoresis assay
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Measuring DNA Damage Using Comet Assay
2
Comet Assay for DNA Damage Assessment
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DMS 114 cells were plated at 500 K cells per well in a six‐well plate, and cells were treated with ±1 nM lurbinectedin and ±2 μM berzosertib for 6 h and collected. Comet assays were performed using the Comet Assay Single Cell Gel Electrophoresis Assay (Trevigen, Gaithersburg, MD) according to manufacturer's instructions. Images were captured using BioSpa Live Cell Analysis System (Biotek) and comet tail length was calculated using OpenComet (https://cometbio.org/ ), a plugin for the image processing program ImageJ.
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