Comet Assay Lab Report

The comet assay, also known as single-cell gel electrophoresis (SCGE), is a standard lab technique that can measure deoxyribonucleic acid (DNA) strand breaks in eukaryotic cells. This technique originated in 1984 with the purpose of quantifying the amount of DNA damage in a cell (3). Originally the test was run using neutral conditions, but this only allowed for detection of double stranded DNA breaks. In 1988 the technique was adapted to use alkaline conditions that made it more sensitive and allowed for analysis of double stranded and single stranded breaks and alkali-labile sites (apurinic and apyrimidinic). The most common use of this test is to analyze white blood cells or lymphocytes in human biomonitory studies, but comet assay can also be used in toxicology, pharmacy, genotoxicity, and genetic disorder diagnosis practices (4). Comet assay has evolved into a commonly used technique because it is simple, cost-effective, has numerous applications, and has fairly rapid results (3). In order to better understand the comet assay, its procedure, analysis, application, and limitations will be examined. In order to perform a comet assay, two days are normally needed. On the first day the cell sample is collected and a microscope slide is prepared be embedding the cell in agrose (1,4). The cell is then …show more content…
This technique answers many questions pertaining to background levels of DNA damage in normal and abnormal cells, variations in repair, regulation of DNA repair, and molecular levels in nucleus. The procedure is fairly simple and can be performed easily by placing a cell on a agrose slide, performing cell lysis, electrophoresis, neutralization, fixation, staining, and microscopy analysis. The comet assay is an important and easy technique, which can be used to determine the about of double stranded and single stranded DNA breaks in a