What is the best buffer for preservation of cells in vitro: a standardization for gill cells in order to use in the Comet Assay
Abstract
The Comet Assay is a rapid and sensitive method for detection of breaks in the DNA strand into individual cells, commonly used at ecotoxicology. At this time, the method has been refined, but not yet fully standardized and variations on protocol are common. When environmental reviews are performed, the large number of samples required and the conditions of handling during transportation of samples to the laboratory are frequent problems. Until now, however, no truly effective method for samples preservation for this test was described. Therefore, we tested three stock solutions: fetal bovine serum (FBS), phosphate buffered saline (PBS) e Tris. Gill cells of Rhamdia quelen contaminated with insecticide Fipronil were maintained in these solutions, for 0, 24 and 48 hours to perform Comet Assay. It was found that fetal bovine serum was the solution that best preserved the DNA integrity of gill cells, followed by Tris and finally by PBS. We concluded in this study that fetal bovine serum is the solution that best conserve the gill cells and genetic material of R. quelen for a time until 48 hours in the absence of light and at 4ºC.
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