Full Description
This volume presents state-of-the art methods for the synthesis, design, assembly, post synthesis processing, and application of synthetic DNA to modern biotechnology. Chapters are divided into three general sections focusing on protocols for the computational design of synthetic DNA sequences, the synthesis, assembly and cloning of synthetic DNA, and post-synthesis error reduction strategies. 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.
Authoritative and cutting-edge, Synthetic DNA: Methods and Protocols aims to help researchers further their research on manipulate DNA sequences.
Contents
A Guide to Using STITCHER for Overlapping Assembly PCR Applications.- Synthetic Gene Design Using Codon Optimization On-Line (COOL).- Shuffle Optimizer: A Program to Optimize DNA Shuffling for Protein Engineering.- Simple Cloning by Prolonged Overlap Extension PCR with Application to the Preparation of Large-Size Random Gene Mutagenesis Library in Escherichia coli.- SpeedyGenes: Exploiting an Improved Gene Synthesis Method for the Efficient Production of Error-corrected, Synthetic Protein Libraries for Directed Evolution.- BASIC - A Simple and Accurate Modular DNA Assembly Method.- Enzymatic Synthesis of Single Stranded Clonal Pure Oligonucleotides.- Rapid Assembly of DNA via Ligase Cycling Reaction (LCR).- PaperClip: A Simple Method for Flexible Multi-part DNA Assembly.- The Polymerase Step Reaction (PSR) Method for Gene and Library Synthesis.- Clonetegration Using OSIP Plasmids - One Step DNA Assembly and Site Specific Genomic Integration in Bacteria.- Generation of DNA Constructs Using the Golden GATEway Cloning Method.- Gene Deletion by Synthesis in Yeast.- Efficient Assembly of DNA Using Yeast Homologous Recombination (YHR).- Simultaneous Removal of Multiple DNA Segments by Polymerase Chain Reactions.- Rapid Construction of Recombinant Plasmids by QuickStep-Cloning.- Immobilized MutS-mediated Error Removal of Microchip-synthesized DNA.- Selection of Error-less Synthetic Genes in Yeast.
