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Synthetic DNA

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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 18: Selection of Error-Less Synthetic Genes in Yeast.
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Chapter title
Selection of Error-Less Synthetic Genes in Yeast.
Chapter number 18
Book title
Synthetic DNA
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6343-0_18
Pubmed ID
Book ISBNs
978-1-4939-6341-6, 978-1-4939-6343-0
Authors

Hisashi Hoshida, Tohru Yarimizu, Rinji Akada

Editors

Randall A. Hughes

Abstract

Conventional gene synthesis is usually accompanied by sequence errors, which are often deletions derived from chemically synthesized oligonucleotides. Such deletions lead to frame shifts and mostly result in premature translational terminations. Therefore, in-frame fusion of a marker gene to the downstream of a synthetic gene is an effective strategy to select for frame-shift-free synthetic genes. Functional expression of fused marker genes indicates that synthetic genes are translated without premature termination, i.e., error-less synthetic genes. A recently developed nonhomologous end joining (NHEJ)-mediated DNA cloning method in the yeast Kluyveromyces marxianus is suitable for the selection of frame-shift-free synthetic genes. Transformation and NHEJ-mediated in-frame joining of a synthetic gene with a selection marker gene enables colony formation of only the yeast cells containing synthetic genes without premature termination. This method increased selection frequency of error-less synthetic genes by 3- to 12-fold.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 6 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
China 1 17%
Unknown 5 83%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 2 33%
Researcher 1 17%
Other 1 17%
Student > Master 1 17%
Unknown 1 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 3 50%
Agricultural and Biological Sciences 2 33%
Unknown 1 17%