SYBR Gold and SYBR Green II are not mutagenic in the Ames test

Favorable photo-physical properties and high affinity to nucleic acids make new fluorescent cyanine dyes of the SYBR-type particularly useful for DNA and RNA visualization. The growing popularity of SYBR-type dyes is also explained by the fact that removal of the dye from the nucleic acids by ethanol precipitation is more efficient and less time-consuming than the phenol–chloroform extraction applied for the widely used phenanthridine DNA stain, ethidium bromide. To evaluate the safety of nucleic acid staining by SYBR Gold and SYBR Green II we compared the mutagenicity of these compounds, with characteristics corresponding to those of ethidium bromide, by use of the Salmonella/mammalian microsome reverse-mutation assays (Ames test). SYBR Green II and SYBR Gold did not show mutagenicity either in frame-shift or in base-substitution indicator strains, TA98 and TA100, respectively. These results were observed both in the presence and in the absence of supernatant from a rat-liver homogenate S9. Mutagenicity of these stains was not observed although their toxic concentration was reached. Toxic effects of SYBR Green II and SYBR Gold were seen approximately at the same molar concentrations as reported previously for SYBR Green I. As expected, ethidium bromide revealed strong mutagenicity with a maximum increase of 60-fold above the vehicle controls in the frame-shift indicator strain TA98 in the presence of rat-liver S9 extract. Thus, SYBR Gold and SYBR Green II do not show mutagenicity in our tests, even at toxic doses, and these DNA stains represent safer alternatives to ethidium bromide for nucleic acid visualization.