Titlebook: Directed Evolution Library Creation; Methods and Protocol Elizabeth M.J. Gillam,Janine N. Copp,David Ackerle Book 2014Latest edition Spring

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Methods: Named Message Sequencescols. SeSaM is advantageous with respect to (1) elimination of mutagenic “hot spots”, (2) increase in frequency of subsequent nucleotide substitutions, (3) control over the mutational bias through the utilization of universal base analogs, and, consequently, (4) the prospect of generating transversi

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https://doi.org/10.1007/978-1-4302-0037-6e amino acid positions from the chosen enzyme are assigned to multi-residue sites (i.e., groups of amino acids or “multisites”). Then, the residues in each multisite are mutated with a user-defined randomization scheme to all canonical amino acids or a reduced amino acid alphabet. Subsequently, the

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Installation and Creating a Robotonal diversities while avoiding high loads of deleterious mutations. Here we describe a simple protocol for incorporating synthetic oligonucleotides that encode designed, site-specific mutations by assembly PCR. This protocol enables a researcher to “hedge the bets,” namely, to explore a large numbe