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The pocket manual of directed evolution: Tips and tricks
Maté D, González-Pérez D, Mateljak I, Gómez de Santos A, Vicente I, Alcalde M

Biotechnology of Microbial Enzymes: Production, Biocatalysis and Industrial Applications. Elsevier, Amsterdam: 185-214

Directed evolution is a robust approach to tailor enzymes with improved attributes. One round of laboratory evolution comprises three essential steps: (1) the generation of diversity by random mutagenesis and/or DNA recombination of the parental genes, (2) cloning and functional expression of the mutant library in a suitable host, and (3) selection or screening for the desired feature. This pocket manual of directed evolution provides an overview of the current methods and protocols used for steps (1) to (3), while giving clever tips and tricks to circumvent common hurdles that appear during creation of the mutant library and its exploration. The new research trends in the evolutionary field (genetic drift, circular permutation, de novo enzyme design, metagenomics, and ancestral resurrection) are also mentioned briefly.

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