Comet assay

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Comet Assay

The Comet Assay (pronounced: /ˈkɒmɪt əˈseɪ/), also known as single cell gel electrophoresis (SCGE), is a sensitive and versatile technique for the measurement of DNA damage in individual cells.

Etymology

The term "Comet Assay" is derived from the characteristic "comet-like" appearance of DNA migration patterns in the gel. The "head" of the comet consists of undamaged DNA, while the "tail" contains damaged or broken pieces of DNA.

Procedure

The Comet Assay begins with the isolation of single cells, which are then embedded in a thin layer of agarose gel on a microscope slide. The cells are lysed to remove proteins and other cellular components, leaving behind the DNA. The slide is then subjected to electrophoresis, which causes the negatively charged DNA to migrate towards the positive electrode. The extent and direction of this migration is determined by the amount of DNA damage present in the cells.

Applications

The Comet Assay is widely used in genotoxicity testing, biomonitoring studies, and human epidemiology studies. It can detect a wide range of DNA damage types, including DNA strand breaks, oxidative DNA damage, and DNA crosslinks.

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