About
Comet Assays
What is a Comet
Assay ?
The comet assay,
also called the 'Single Cell Gel Assay', is the technique to detect
DNA damage and repair at the level of single cells. This is one of
the advanced technique in the oncology. This technique was developed
by Swedish researchers Östling & Johansson in 1984. This technique
was later modified by N.P. Singh et al., in 1988 as Alkaline Comet
Assay.
'Comet Assay' is one of the most popular tests of DNA damage
detection (e.g., single- and double-strand breaks,
oxidative-induced base damage, and DNA-DNA/DNA-protein cross linking
) by electrophoresis, developed in recent years.
Advantages Of Comet
Assay :
-
Very high sensitivity to detect DNA damage
-
Rapid and easy to handle
-
Little amount of cell samples needed
-
Applied to most eukaryotic cells
Methodology
:
Comet Assay involves the following steps :
1. Slide preparation : Mixing of cells with low melting agarose,
and spread over glass microscope slides
2. Lysis: lysis of cell membrane and other proteins
3. Unwinding of DNA
4. Electrophoresis
5. Neutralization
6. Staining and scoring
Applications
:
-
Genotoxicity
Studies
-
DNA Repair
-
Apoptosis
-
Visual
quantification of DNA double strand breaks in bacteria
-
Free Radical
Biology
-
Nutritional Toxicology
-
Biomonitoring
-
Clinical Applications
-
Cell cycle analysis
