DNA Damage Assay HCT 116

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 2 matching solutions for this experiment

CometAssay Single Cell Gel Electrophoresis Assay

Bio-Techne

Upstream tips	Protocol tips	Downstream tips
Seed 1–2 × 10^4 cells/ml	For neutral comet assay, the slides were 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 were immersed in 70 % EtOH for 5 min after electrophoresis then incubated with SYBR®Gold for 30 min.	For statistical analysis use OpenComet, an ImageJ plugin

Upstream tips
Seed 1–2 × 10^4 cells/ml
Protocol tips
For neutral comet assay, the slides were 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 were immersed in 70 % EtOH for 5 min after electrophoresis then incubated with SYBR®Gold for 30 min.
Downstream tips
For statistical analysis use OpenComet, an ImageJ plugin

Manufacturer protocol 1 Relevant paper

CometAssay Single Cell Gel Electrophoresis Assay-Silver

Bio-Techne

Upstream tips	Protocol tips	Downstream tips
Suspend cell pellet at 3 x 10^5 cells/mL in 10% (v/v) DMSO, 40% (v/v) medium, 50% (v/v) fetal bovine serum.		Use the OPENCOMET plugin for calculating the tail moment of a comet.

Upstream tips
Suspend cell pellet at 3 x 10^5 cells/mL in 10% (v/v) DMSO, 40% (v/v) medium, 50% (v/v) fetal bovine serum.
Downstream tips
Use the OPENCOMET plugin for calculating the tail moment of a comet.

Manufacturer protocol Publication protocol 1 Relevant paper

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