MADD-seq yeast 2023
Our genome is exposed to a wide variety of DNA-damaging agents. If left unrepaired, this damage can be fixed into mutations that promote carcinogenesis and the development of genetically inherited diseases. As a result, it is crucial that we can detect DNA damage and mutations with exquisite sensitivity. Here, we describe a modified version of double barcoding sequencing technology termed Mutation And DNA Damage Detection-seq (MADDDseq) that can detect DNA damage and mutations simultaneously, with a single assay. To demonstrate the utility of MADDD-seq as a multifunctional detection tool, we treated yeast cells with a DNA-damaging agent and tracked the presence of DNA damage and mutations over a 24-hour timespan. These experiments allowed us to identify thousands of adducts and mutations in a single sequencing run and expose the kinetics of DNA repair and mutagenesis in remarkable detail.