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Synthetic DNA

Overview of attention for book
Cover of 'Synthetic DNA'

Table of Contents

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    Book Overview
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    Chapter 1 A Guide to Using STITCHER for Overlapping Assembly PCR Applications.
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    Chapter 2 Synthetic Gene Design Using Codon Optimization On-Line (COOL).
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    Chapter 3 Shuffle Optimizer: A Program to Optimize DNA Shuffling for Protein Engineering.
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    Chapter 4 Simple Cloning by Prolonged Overlap Extension-PCR with Application to the Preparation of Large-Size Random Gene Mutagenesis Library in Escherichia coli.
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    Chapter 5 Synthetic DNA
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    Chapter 6 BASIC: A Simple and Accurate Modular DNA Assembly Method.
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    Chapter 7 Enzymatic Synthesis of Single-Stranded Clonal Pure Oligonucleotides.
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    Chapter 8 Rapid Assembly of DNA via Ligase Cycling Reaction (LCR).
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    Chapter 9 PaperClip: A Simple Method for Flexible Multi-Part DNA Assembly.
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    Chapter 10 The Polymerase Step Reaction (PSR) Method for Gene and Library Synthesis.
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    Chapter 11 Clonetegration Using OSIP Plasmids: One-Step DNA Assembly and Site-Specific Genomic Integration in Bacteria.
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    Chapter 12 Generation of DNA Constructs Using the Golden GATEway Cloning Method.
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    Chapter 13 Synthetic DNA
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    Chapter 14 Efficient Assembly of DNA Using Yeast Homologous Recombination (YHR).
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    Chapter 15 Simultaneous Removal of Multiple DNA Segments by Polymerase Chain Reactions.
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    Chapter 16 Synthetic DNA
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    Chapter 17 Immobilized MutS-Mediated Error Removal of Microchip-Synthesized DNA.
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    Chapter 18 Selection of Error-Less Synthetic Genes in Yeast.
Attention for Chapter 4: Simple Cloning by Prolonged Overlap Extension-PCR with Application to the Preparation of Large-Size Random Gene Mutagenesis Library in Escherichia coli.
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Chapter title
Simple Cloning by Prolonged Overlap Extension-PCR with Application to the Preparation of Large-Size Random Gene Mutagenesis Library in Escherichia coli.
Chapter number 4
Book title
Synthetic DNA
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6343-0_4
Pubmed ID
Book ISBNs
978-1-4939-6341-6, 978-1-4939-6343-0
Authors

Chao Zhong, Chun You, Ping Wei, Yi-Heng Percival Zhang, Zhong, Chao, You, Chun, Wei, Ping, Zhang, Yi-Heng Percival

Editors

Randall A. Hughes

Abstract

We developed a simple method (simple cloning) for subcloning DNA fragments into any location of a targeted vector without the need of restriction enzyme, ligase, exonuclease, or recombinase in Escherichia coli. This technology can be applied to common E. coli hosts (e.g., DH5α, JM109, TOP10, BL21(DE3)). The protocol includes three steps: (1) generate DNA insert and linear vector backbone by regular high-fidelity PCR, where these two DNA fragments contain 3' and 5' overlapping termini; (2) generate DNA multimers based on these two DNA fragments by using prolonged overlap extension-PCR (POE-PCR) without primers added; and (3) transform POE-PCR product to competent Escherichia coli cells directly, yielding the desired plasmid. Simple cloning provides a new cloning method with great simplicity and flexibility. Furthermore, this new method can be modified for the preparation of a large-size mutant library for directed evolution in E. coli. Using this method, it is very easy to generate a mutant library with a size of more than 10(7) per 50 μL of the POE-PCR product within 1 day.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 44 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 1 2%
Unknown 43 98%

Demographic breakdown

Readers by professional status Count As %
Researcher 9 20%
Student > Ph. D. Student 8 18%
Student > Master 6 14%
Student > Bachelor 5 11%
Student > Doctoral Student 2 5%
Other 6 14%
Unknown 8 18%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 13 30%
Agricultural and Biological Sciences 7 16%
Chemistry 4 9%
Immunology and Microbiology 3 7%
Chemical Engineering 2 5%
Other 6 14%
Unknown 9 20%