DNA Damage Assay U-2 OS

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.

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Found 3 matching solutions for this experiment

CometAssay Single Cell Gel Electrophoresis Assay

Bio-Techne

Upstream tips	Protocol tips	Downstream tips
Seed 1,00,000cells/ml	For neutral comet assay, the slides are to be immersed in 1 × TBE buffer for electrophoresis at 1 volt/cm for 30 min at room temperature. For both alkaline and neutral comet assays, the slides can be immersed in 70 % EtOH for 5 min after electrophoresis then incubated with SYBR®Gold for 30 min.	The images can be visualized under BioTek Cytation 5 cell imaging reader. Statistical analysis was performed by OpenComet, an ImageJ plugin

Upstream tips
Seed 1,00,000cells/ml
Protocol tips
For neutral comet assay, the slides are to be immersed in 1 × TBE buffer for electrophoresis at 1 volt/cm for 30 min at room temperature. For both alkaline and neutral comet assays, the slides can be immersed in 70 % EtOH for 5 min after electrophoresis then incubated with SYBR®Gold for 30 min.
Downstream tips
The images can be visualized under BioTek Cytation 5 cell imaging reader. Statistical analysis was performed by OpenComet, an ImageJ plugin

Manufacturer protocol Publication protocol 1 Relevant paper

APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
Seed 2 × 10^5 cells/ml/well

Upstream tips
Seed 2 × 10^5 cells/ml/well

Manufacturer protocol Publication protocol 1 Relevant paper

CometAssay Electrophoresis System II

Bio-Techne

Upstream tips	Protocol tips	Downstream tips
Seed 50,000 cells per well in a 96-well plate		Comets can be imaged by using the LionHeart FX automated microscope (Biotek) and the Gen5 Image Prime software to construct image montages that were analyzed using TriTek CometScore 2.0.0.38.

Upstream tips
Seed 50,000 cells per well in a 96-well plate
Downstream tips
Comets can be imaged by using the LionHeart FX automated microscope (Biotek) and the Gen5 Image Prime software to construct image montages that were analyzed using TriTek CometScore 2.0.0.38.

Manufacturer protocol Publication protocol 1 Relevant paper

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