Protocol for adaptive laboratory evolution of S. cerevisiae by PEG/LiAc transformation and sequencing

Gene deletions perturb cellular homeostasis, affecting gene expression and phenotypes. Here, we present a protocol for serial transfer experiments in Saccharomyces cerevisiae strains following targeted gene knockouts. We describe steps for gene deletion, serial passage optimization, and growth kinetics analysis. We then detail procedures for genomic and transcriptomic profiling. This protocol enables the investigation of changes in yeast strains resulting from gene loss. For complete details on the use and execution of this protocol, please refer to Jiang et al.¹.