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This volume provides a comprehensive collection of DNA assembly protocols that will prove useful for any researcher interested in molecular cloning, synthetic biology, or DNA manipulation. Chapters will guide readers through computational tools to design and track the construction of DNA assemblies, workflows that enable high-throughput assembly of DNA constructs, standardized toolkits and protocols for DNA assembly, and combinatorial solutions that enable the construction and optimization of entire metabolic pathways. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, DNA Cloning and Assembly: Methods and Protocols aims to ensure successful results in the further study of this vital field.
List of contents
An Integrated Computer-Aided Design and Manufacturing Workflow for Synthetic Biology.- Combinatorial-Hierarchical DNA Library Design Using the TeselaGen DESIGN Module with j5.- The AssemblX Toolkit for Reliable and User-friendly Multi-gene Assemblies.- High-Throughput PCR for DNA Part Generation.- High-Throughput DNA Assembly Using Yeast Homologous Recombination.- Highly-Multiplexed, Semi-Automated Nextera Next Generation Sequencing (NGS) Library Preparation.- Generation of MoClo Standard Parts Using Golden Gate Cloning.- Assembly of Multigene Constructs using the Modular Cloning System MoClo.- Hierarchical Modular DNA Assembly using MetClo.- PaperClip DNA Assembly: Reduce, Reuse, Recycle.- Phytobricks: Manual and Automated Assembly of Constructs for Engineering Plants.- Mobius Assembly.- Design and Implementation of Multi-Protein Expression Constructs and Combinatorial Libraries using Start-Stop Assembly.- BASIC - A Simple and Accurate Modular DNA Assembly Method.- Joint Universal Modular Plasmids: A Flexible Platform for Golden Gate Assembly in Any Microbial Host.- A Step-by-Step Protocol for COMPASS, a Synthetic Biology Tool for Combinatorial Gene Assembly.- SCRaMbLE-in: A Fast and Efficient Method to Diversify and Improve the Yields of Heterologous Pathways in Synthetic Yeast.
