What is a comet assay
16 April 2022
The Comet Assay, commonly known as single-cell gel electrophoresis (SCGE), is a versatile, simple, and adaptable method for detecting DNA damage and repairing single cells. It is based on the ability of negatively charged loops/fragments of DNA to be pushed through an agarose gel by an electric field, giving the appearance of a ‘comet.’
The Comet Assay has grown in popularity over the last two decades, and it is now the most widely used assay for assessing DNA damage and repair. It combines the single-cell approach of cytogenetic assays with the simplicity of biochemical techniques for identifying single-strand DNA single-strand breaks, including strand breaks, partial excision repair sites, and alkali-labile sites (ALS) cross-linking. (strand breaks and incomplete excision repair sites).
Advantages of using comet Assay
The Comet Assay has several advantages, including
(1) sensitivity for detecting low levels of damage
(2) use of any monodispersed cell population, both proliferating and nonproliferating
(3) single-cell data collection for
(4) the demand for a minimal number of cells per sample,
(5) low cost, speed, and convenience of application
(6) the flexibility to use fresh or frozen samples.
The assay’s popularity stems from the fact that it may be used with any eukaryote cell that can be collected as a single cell suspension, such as cells separated from the blood or homogenized tissue biopsies, buccal cells, whole blood, and cultured cells. However, some cell types, such as plant cells and sperm cells, necessitate a few tweaks to the standard technique. In general, for most reasons, well-characterized cell lines or primary cells (e.g., peripheral blood mononuclear cells) utilized in traditional genetic toxicology testing tests are preferable.
Cost of comet Assay
The comet assay is a low-cost and highly sensitive approach for detecting sperm DNA damage. The comet assay’s measurement of sperm DNA damage is closely linked to other direct methods like the TUNEL assay (Shamsi et al., 2008). DNA damage can be measured in cell types, including sperm, using the comet assay. The operation can be carried out with only a few cells. Because the data is measured with software, it is more exact and less susceptible to experimental bias. The comet assay measures DNA damage linked to male infertility and ART outcomes .