Applicability of Environmental Microorganisms to Alkaline Single Cell Gel Electrophoresis (Comet) Assay

Abstract

Possibilities of the alkaline single cell gel electrophoresis (comet) assay were experimentally investigated, using 5 kinds of environmental microorganisms i.e. Euglena gracilis, Chlamydomonas pulsatilla, Saccharomyces cerevisiae, Monas guttula and Tetraphymena pyriformis. Consequently, applicability of E.gracilis to comet assay was found and its most suitable modes of operation were determined to be cell lysis for 5 min by 0.01%-sodium dodecyl sulfate (SDS) solution, unwinding for 20 min and electrophoresis (current: 300mA, voltage: 25 V) for 20 min, respectively. Tail length and Tail moment have been confirmed to be important as evaluation indices of comet assay through the experiments of γ-irradiation.