Genetic screens and directed evolution can be performed in microdroplets to achieve high-throughput sample processing and analysis. In this workflow, mutant enzyme libraries generated via error-prone PCR are encapsulated in droplets and incubated. A fluorescent substrate allows active variants to be sorted and recovered for next-generation sequencing. Data analysis reveals the relative entropy of each residue in the protein structure, indicating which are critical to protein function. Using these principles, functional enzyme screens can be designed to maximize, for example, enzymatic production of a natural product.
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