What is tail moment in comet assay?
What is tail moment in comet assay?
The tail moment is defined as the product of the tail length and the fraction of total DNA in the tail (Tail moment=tail length x % of DNA in the tail). This is calculated automatically by the computer software system as an average for the 50 cells selected for measurement.
What is DNA tail called as?
If DNA damage has occurred the electrophoresed DNA fragments appear as a diffuse tail behind the nucleus known as a “comet”. Many computer-aided analysis systems are currently in use to quantify the amount of DNA damage that is represented by a comet image.
How do you calculate Olive tail moment?
OTM was used for DNA fragmentation (damage) quantification, and was calculated as: OTM = (Tail. mean – Head. mean) × (Tail %DNA)/100.
How can you detect DNA damage?
Breaks in DNA reduce the molecular weight of a single DNA strand, and this may be caused by physical, chemical or enzymatic reagents (6). DNA breaks and lesions may be detected by PCR or using agarose gel electrophoresis (7). PCR is one of the most frequently used techniques for detecting DNA damage (7).
How do you do a comet assay?
Comet assay utilizes single cells to measure DNA damage. First, cells are embedded into agarose and then placed onto a slide. The slide is then immersed into lysis solution to break open the cell membrane. After the cells are lysed, DNA is denatured under neutral or alkaline conditions and run through electrophoresis.
What is neutral comet assay?
Comet assay under neutral conditions allows the detection of DNA double-strand breaks (DSB), considered to be the biologically relevant radiation-induced lesion.
What is comet assay technique?
The Comet Assay, also called single cell gel electrophoresis (SCGE), is a sensitive and rapid technique for quantifying and analyzing DNA damage in individual cells. As such, this is one of the techniques used in the area of cancer research for the evaluation of genotoxicity and effectiveness of chemoprevention.
What is single cell gel electrophoresis?
The Comet Assay, also called single cell gel electrophoresis (SCGE), is a sensitive and rapid technique for quantifying and analyzing DNA damage in individual cells. Individual cells are embedded in a thin agarose gel on a microscope slide. All cellular proteins are then removed from the cells by lysing.
How does comet assay help in DNA damage detection?
The comet assay (single-cell gel electrophoresis) is a simple method for measuring deoxyribonucleic acid (DNA) strand breaks in eukaryotic cells. DNA repair can be monitored by incubating cells after treatment with damaging agent and measuring the damage remaining at intervals.
How does the comet assay work?
The comet assay is based upon the movement of nuclear DNA through an agarose gel when an electrical field is applied. Simply put, the amount of DNA which leaves the nucleus is a measure of the amount of DNA damage to the cell. The brighter and longer the DNA tail, the higher the level of damage.
