製剤科学とバイオアベイラビリティのジャーナル

Single Cell Gel Electrophoresis and Its Applications in Different Fields

Abstract

Ali Ahmed

The Single Cell Gel Electrophoresis assay (SCGE) is a better technique for the detection of DNA damage and DNA repair in eukaryotes. First time comet assay technique was designed by Ostling and Johanson in 1984 and then it was modified by Singh et al. in 1988. There are four main steps in comet assay. These are:
1. Encapsulation of cell in agarose having low melting point.
2. Lysis of cells.
3. Gel electrophoresis and forth is staining and visualization.
This technique is also known as comet assay because DNA movement pattern in gel electrophoresis similar to a comet. The comet assay is use for the measurement of DNA strand breaks in eukaryotes. In this technique cell suspended in agarose on a slide and lysed with detergent to form nucleoid contain supercoiled DNA attached with nuclear matrix. A comet which was made in electrophoresis detected by fluorescence microscopy, manual or mechanical methods use for scoring but now a day’s mechanical methods are very useful. Comet assay have simplicity, sensitivity, speed and economical use. In this review we discuss its protocol and its applications in different fields.