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DNA Cloning and Assembly Methods

Overview of attention for book
Cover of 'DNA Cloning and Assembly Methods'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 BioBrick Assembly Standards and Techniques and Associated Software Tools.
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    Chapter 2 Plasmid Construction by SLIC or Sequence and Ligation-Independent Cloning
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    Chapter 3 Quick and Clean Cloning
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    Chapter 4 Hierarchical ligation-independent assembly of PCR fragments.
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    Chapter 5 USER-Derived Cloning Methods and Their Primer Design
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    Chapter 6 Application of the Restriction-Free (RF) Cloning for Multicomponents Assembly
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    Chapter 7 A Single-Tube Assembly of DNA Using the Transfer-PCR (TPCR) Platform.
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    Chapter 8 Circular polymerase extension cloning.
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    Chapter 9 Golden gate cloning.
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    Chapter 10 Design and Construction of Multigenic Constructs for Plant Biotechnology Using the GoldenBraid Cloning Strategy
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    Chapter 11 FX Cloning: A Simple and Robust High-Throughput Cloning Method for Protein Expression
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    Chapter 12 Minimum GC-Rich Sequences for Overlap Extension PCR and Primer Annealing
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    Chapter 13 Simple Cloning and DNA Assembly in Escherichia coli by Prolonged Overlap Extension PCR
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    Chapter 14 Combinatorial Assembly of Clone Libraries Using Site-Specific Recombination
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    Chapter 15 Application of In-Fusion™ Cloning for the Parallel Construction of E. coli Expression Vectors
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    Chapter 16 Seamless Ligation Cloning Extract (SLiCE) Cloning Method.
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    Chapter 17 j5 DNA Assembly Design Automation
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    Chapter 18 FastPCR Software for PCR, In Silico PCR, and Oligonucleotide Assembly and Analysis.
Attention for Chapter 13: Simple Cloning and DNA Assembly in Escherichia coli by Prolonged Overlap Extension PCR
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Chapter title
Simple Cloning and DNA Assembly in Escherichia coli by Prolonged Overlap Extension PCR
Chapter number 13
Book title
DNA Cloning and Assembly Methods
Published in
Methods in molecular biology, January 2014
DOI 10.1007/978-1-62703-764-8_13
Pubmed ID
Book ISBNs
978-1-62703-763-1, 978-1-62703-764-8
Authors

Chun You, Y.-H. Percival Zhang

Abstract

We developed a simple method (Simple Cloning) for subcloning one, two, or three DNA fragments into any location of a targeted vector without the need for restriction enzyme, ligase, exonuclease, or recombinase. This cloning technology can be applied to a few common Escherichia coli hosts (e.g., BL21(DE3), DH5α, JM109, TOP10). The protocol includes three steps: (a) linear DNA fragments (i.e., the insert DNA and the vector backbone) with two overlap ends were generated by regular high-fidelity PCR, (b) the DNA multimers were generated based on these equimolar DNA templates by using prolonged overlap extension PCR (POE-PCR) without primers added, and (c) the POE-PCR product was transformed to E. coli strains directly. Because positive colony efficiencies are very high, it is not necessary to identify desired clones by using colony PCR. Simple Cloning provides a new cloning and DNA assembly method with great simplicity and flexibility.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 30 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
France 1 3%
Unknown 29 97%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 10 33%
Researcher 8 27%
Lecturer 2 7%
Professor 1 3%
Student > Master 1 3%
Other 1 3%
Unknown 7 23%
Readers by discipline Count As %
Agricultural and Biological Sciences 13 43%
Biochemistry, Genetics and Molecular Biology 4 13%
Materials Science 1 3%
Unknown 12 40%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 15 July 2014.
All research outputs
#20,233,066
of 22,758,963 outputs
Outputs from Methods in molecular biology
#9,865
of 13,089 outputs
Outputs of similar age
#264,801
of 305,266 outputs
Outputs of similar age from Methods in molecular biology
#405
of 597 outputs
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