Golden Gate Cloning : Methods and Protocols / edited by Daniel Schindler.

Format:

Book

Contributor:

Schindler, Daniel E., Editor.

Series:

Methods in Molecular Biology, 1940-6029 ; 2850

Language:

English

Subjects (All):

Genetics.

Local Subjects:

Genetics.

Physical Description:

1 online resource (XIV, 503 p. 93 illus., 90 illus. in color.)

Edition:

1st ed. 2025.

Place of Publication:

New York, NY : Springer US : Imprint: Humana, 2025.

Summary:

This volume provides an overview of the latest protocols that cover the full range of in silico, in vitro, and in vivo approaches to help researchers plan, design, and perform Golden Gate cloning experiments. The chapters in this book cover topics such as Golden Gate cloning of standardized parts; modular DNA construct design for high-throughput Golden Gate Assembly; Golden Gate cloning of Synthetic CRISPR RNA spacer sequences; Golden Gate cloning in Actinobacteria; and YeastFab cloning of toxic genes and protein expression optimization in yeast. 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 protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Golden Gate Cloning: Methods and Protocols is a valuable resource for all researchers interested in learning more about this important and developing field. Chapter 3 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

Golden Gate Cloning of MoClo Standard Parts

Assembly of Multigene Constructs using the Modular Cloning System MoClo

Selection of Fusion-Site Overhang Sets for High Fidelity and High Complexity Golden Gate Assembly

Modular DNA Construct Design for High-Throughput Golden Gate Assembly

Using ApE for In Silico Golden Gate Cloning

Standardized Golden Gate Assembly Metadata Representation using SBOL

Use of a Golden Gate Plasmid Set Enabling Scarless MoClo-Compatible Transcription Unit Assembly

Biofoundry-Assisted Golden Gate Cloning with AssemblyTron

Automation and Miniaturization of Golden Gate DNA Assembly Reactions using Acoustic Dispensers

Validation of Golden Gate Assemblies using Highly Multiplexed Nanopore Amplicon Sequencing

Modular Golden Gate Assembly of Linear DNA Templates for Cell-Free Prototyping

Construction of Small Genes with Repetitive Elements using Oligo Extension and Golden Gate Assembly

Simplifying Recombinant Protein Production: Combining Golden Gate Cloning with a Standardized Protein Purification Scheme

Generating Site Saturation Mutagenesis Libraries and Transferring Them to Broad Host-Range Plasmids using Golden Gate Cloning

Combining Oligo Pools and Golden Gate Cloning to Create Protein Variant Libraries or Guide RNA Libraries for CRISPR Applications

Golden Gate Cloning of Synthetic CRISPR RNA Spacer Sequences

Golden Gate Cloning of Expression Plasmids for Synthetic Small RNAs in Bacteria

Utilizing Golden Gate Assembly to Streamline CRISPR-Cas/NgTET-Based Phage Mutagenesis

Golden Gate Assisted Gene Doctoring for Streamlined and Efficient Recombineering in Bacteria

Protocol for NT-CRISPR, A Method for Efficient Genome Engineering in Vibrio natriegens

Golden Gate Cloning in Actinobacteria: Opportunities and Challenges

Golden Gate Assembly of Transcriptional Unit Libraries into a Rearrangeable Gene Cluster

Golden Gate Cloning for Efficient Biosynthesis of Lycopene in Synthetic Yeast

YeastFab Cloning of Toxic Genes and Protein Expression Optimization in Yeast

Golden Gate Cloning for the Standardized Assembly of Gene Elements with Modular Cloning in Chlamydomonas

Assembling DNA Plasmids with the Multi-Kingdom (MK) Cloning System

Advancements in Golden Gate Cloning: A Comprehensive Review.

ISBN:

9781071642207

1071642200