A MODIFIED PROTOCOL OF COMET ASSAY ON HUMAN BUCCAL CELL

摘要

In order to improve the efficiency of comet assay, which can be used in epidemiological research to detect genotoxicity of air pollutants, we undertook this study. Formaldehyde at 50μM was employed as contaminating agent to poison the human buccal cell before comet assay. During the process of comet assay, each pair of contaminated samples was treated with proteinase K of three different concentrations at 0.1mg/ml, 1mg/ml and 10mg/ml, respectively while other steps are kept the same. The result turned out to be that the 1 mg/ml proteinase K could lead to an ideal incubating effect. It was also proved that a 45-minute lysis had almost the same effect as the 120-min one. A further try was the reduction of the enzyme-incubation period to 45 min; the result showed that with the treatment of 1mg/ml proteinase K, a 45-minute lysis and a 45-minute enzyme-incubation as well can achieve a relatively ideal result.