Principle of Assay
This kit is a single cell gel electrophoresis assay (SCGE), rapid and sensitive test for detecting DNA damage in cells. The principle is that after organs or tissues have been suffered from (such as radiation, heavy metals, etc.), single stranded or double stranded DNA is damaged within the cells. The cell is mixed with melted agarose and added on a slide, then the cell membrane is disrupted with lysis solution, the DNA is uncoiled with alkaline solution. Finally, the samples are electrophoresed, under the action of an electric field, the macromolecular DNA of normal cells migrates for a shorter distance. Intact DNA remains in the nucleus. Broken pieces of DNA migrate out of the nucleus, stained with PI dye, and observed under a fluorescence microscope. Intact DNA forms a circular or slightly tailed pattern; DNA fragments form a comet-like pattern. According to the pH value of the electrophoresis buffer, it can be divided into neutral comet assay (pH=8.4) and alkaline comet assay (pH>13). Neutral comet assay is mainly used to detect double-stranded DNA beaks. Alkaline comet assay has a higher sensitivity and can be used to detect single- and double-stranded DNA beaks .
