The comet assay is a technique to measure DNA damage and other DNA alterations in individual cells. The technique aquired its name from the comet-like shape of the DNA of the cells which can be seen under the microscope after the procedure. The technique is also named single cell gel electrophoresis assay, which is more descriptive. The main steps of the procedure are preparation of a cell suspension with the cells of interest, embedding cells in an agarose gel and spreading them on an object slide, lysis of cells to remove non-DNA compounds, exposing the DNA to a neutral or alkaline buffer, subjecting the gels with the DNA to electrophoresis (which forms the "comets"), and measuring the extent of migration and stretching of DNA by microscopical analysis. As many cells are measured (e.g. 100 cells per object slide), the statistical distribution of the individual cell measurements represents the amount of DNA damage/alteration of the cell sample.
From-http://www.comet-assay.de/cometintro.php
No comments:
Post a Comment