Pooled yeast knockout library selection under temporally varying temperature regimes.

Fluctuating conditions may yield adaptations distinct from those selected under monotonic pressure. Prior studies often either use highly controlled microfluidic systems to study a few strains at a time, or tedious bulk techniques for limited perturbation on a pooled population of thousands of strains. eVOLVER can provide both control and throughput, enabling temporal changes of one selective pressure on a large population (>10^7 cells) while holding other culture conditions constant. To demonstrate this, we carried out growth selection experiments on a pooled YKO library (5,149 unique members) under conditions in which a single environmental variable—temperature—was temporally varied.