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The Statistics Of Directed Evolution: From Library Generation To High Throughput Screens, Keith Tyo

Biochemical and Molecular Engineering XXI

Directed evolution has been recognized as a powerful approach to creating enzymes and cells with desirable properties, such as growth on a new substrate, thermotolerance, or even novel reaction mechanisms. The directed evolution approach is to generate diversity through genetic mutations and screen the diversity for mutants that have improved properties compared to the parent. This diversification and screening can be repeated to generate mutants with superior properties.

While the concept is straightforward, the devil is in the details. How should I create diversity (error prone PCR, mutagenic primers, transposons…)? How much diversity do I need (10³, 10 …