Here we demonstrate a simple protocol to create a random mutant library for a given target sequence. We show how this method, which is performed in vivo in Escherichia coli, can be coupled with functional selections to evolve new enzymatic activities.
Troll, C., Alexander, D., Allen, J., Marquette, J., Camps, M. Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli. J. Vis. Exp. (49), e2505, doi:10.3791/2505 (2011).