Adaptive laboratory evolution of antimicrobial resistance in bacteria for genetic and phenotypic analyses

Adaptive laboratory evolution (ALE) of bacteria has the potential to provide many insights like revealing novel mechanisms of resistance and elucidating the impact of drug combinations and concentrations on AMR evolution. Here, we describe a step-by-step ALE protocol for the model bacterium Escherichia coli that can be easily adapted to answer questions related to evolution and genetics of AMR in diverse bacteria. Key issues to consider when designing ALE experiments as well as some downstream mutation mapping analyses are described. For complete details on the use and execution of this protocol, please refer to Patel and Matange (2021)¹ and Matange et al. (2019).².