Directed Evolution: Methods and Protocols

by Andrew Currin (Editor), Neil Swainston (Editor)

Overview

This volume explores the latest techniques used by researchers to study directed evolution (DE) at each stage of the Design-Build-Test-Learn cycle. Chapters in this book cover topics such as designing overlap extension PCR primers for protein mutagenesis; antha-guided automation of Darwin assembly for the construction of bespoke gene libraries; rapid cloning of random mutagenesis libraries using PTO-Quickstep; and DE of glycosyltransferases by a single-cell screening method. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Directed Evolution: Methods and Prools is a valuable resource for scientists and researchers who are interested in learning more about this field and incorporating these studies into new experimental workflows.

Product Details

ISBN-13:	9781071621547
Publisher:	Springer US
Publication date:	06/23/2023
Series:	Methods in Molecular Biology , #2461
Edition description:	1st ed. 2022
Pages:	278
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Designing Overlap Extension PCR Primers for Protein Mutagenesis: A Programmatic Approach.- Recombination of Single Beneficial Substitutions Obtained from Protein Engineering by Computer-Assisted Recombination (CompassR).- Non-Degenerate Saturation Mutagenesis: Library Construction and Analysis via MAX and ProxiMAX Randomization.- Antha-Guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries.- SpeedyGenesXL An Automated, High-Throughput Platform for the Preparation of Bespoke Ultra-Large Variant Libraries for Directed Evolution.- Facile Assembly of Combinatorial Mutagenesis Libraries using Nicking Mutagenesis.- GeneORator: An Efficient Method for the Systematic Mutagenesis of Entire Genes.- Rapid Cloning of Random Mutagenesis Libraries using PTO-QuickStep.- Construction of Strong Promoters by Assembling Sigma Factor Binding Motifs.- Application of Restriction Free (RF) Cloning in Circular Permutation.- Site-Directed Mutagenesis Method Mediated by Cas9.- Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity.- A Screening Method for P450 BM3 Mutant Libraries using Multiplexed Capillary Electrophoresis for Detection of Enzymatically Converted Compounds.- Directed Evolution of Glycosyltransferases by a Single-Cell Ultrahigh-Throughput FACS-Based Screening Method.- Learning Strategies in Protein Directed Evolution.