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This volume details technologies on recombinant DNA and nucleic acid manipulation that underpin much of the biological sciences and instructions on how to conduct them successfully. Chapters guide the reader through the basics of oligonucleotide synthesis and DNA sequencing recombinant DNA plasmid work cell-free experiments and the latest developments in CRISPR approaches to genome modification. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Gene Modification and Nucleic Acid Technologies aims to be the comprehensive guide for life scientists moving into the field of recombinant DNA and nucleic acid manipulation.
Classical Recombinant DNA Cloning
A Sequence- and Ligation-Independent Cloning (SLIC) Procedure for the Insertion of Genes into a Plasmid Vector
Molecular Cloning Using In Vivo DNA Assembly
Assembling Multiple Fragments:​ The Gibson Assembly
TA Cloning Approaches to Cloning DNA with Damaged Ends DNA
PCR-Based Assembly of Gene Sequences by Thermodynamicall​y Balanced Inside-Out (TBIO) Gene Synthesis
Random Mutagenesis by PCR
In Vitro Site Directed Mutagenesis
Xenopus Transgenesis Using the pGateway System
CRISPR/Cas9 Gene Disruption Studies in F0 Xenopus Tadpoles: Understanding Development and Disease in the Frog
A CRISPR/Cas-Based Method for Precise DNA Integration in Xenopus laevis Oocytes Followed by Intracytoplasmic Sperm Injection (ICSI) Fertilization
A Lambda-Exonuclease SELEX Method for Generating Aptamers to Bacterial Targets
Generation of Functional-RNA Arrays by In Vitro Transcription and In Situ RNA Capture for the Detection of RNA-RNA Interactions
Chemical Synthesis of Oligonucelotide Sequences:​ Phosphoramidite Chemistry
Low Throughput Direct Cycle Sequencing of Polymerase Chain Reaction (PCR) Products
Nanopore Sequencing for Mixed Samples
Ethics, Legality, and Safety for Geneticists