In the last half century, various methods such as the Ames test and the micronucleus test have been employed as simple, sensitive, and reliable approaches for evaluation of the mutagenicity of chemicals and drugs ( OECD, 2016 OECD, 2020). We believe that the method presented here will contribute to future opportunities for research on mutagenesis, DNA repair, and cancer. Furthermore, a very small amount of NGS data from selection plates reveals the almost full spectrum of mutations in each specimen and offers us a novel insight into the mechanisms of mutagenesis, despite them being considered already well known. coli grown on titer plates provides mutation frequency and spectrum data, and uncovers obscure mutations that cannot be detected by a conventional supF assay. The NGS data obtained from samples of the indicator E. We demonstrate the performance of our newly developed assay via pilot experiments with ultraviolet (UV) irradiation, the results from which emerge more relevant than expected. In this study, by using a supF shuttle vector and non-SOS-induced Escherichia coli with short-read next-generation sequencing (NGS) technology, we present an advanced method for the study of mutations, which is simple, versatile, and cost-effective. A forward mutagenesis assay using the supF gene has been widely employed for the last several decades in studies addressing mutation frequencies and mutation spectra associated with various intrinsic and environmental mutagens.
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