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E. Coli Plasmid Vectors: Methods and Applications als Buch
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E. Coli Plasmid Vectors: Methods and Applications

Methods and Applications. 'Methods in Molecular Biology'. Auflage 2003. Sprache: Englisch.
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A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, … weiterlesen
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E. Coli Plasmid Vectors: Methods and Applications als Buch

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Titel: E. Coli Plasmid Vectors: Methods and Applications

ISBN: 1588291510
EAN: 9781588291516
Methods and Applications.
'Methods in Molecular Biology'.
Auflage 2003.
Sprache: Englisch.
Herausgegeben von Nicola Casali, Andrew Preston
SPRINGER VERLAG GMBH

3. Juli 2003 - gebunden - 316 Seiten

Beschreibung

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described. Manipulation of recombinant DNA, which is almost exclusively performed using the host E. coli, constitutes one of the fundamental methodologies of molecular biotechnology. In E. coli Plasmid Vectors, experienced bench researchers describe their proven techniques for the manipulation of recombinant plasmids utilizing this popular bacterial host. The authors describe readily reproducible methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Also presented are methods for common downstream applications, such as mutagenesis, expression of recombinant proteins and RNA transcripts, and uses of reporter genes. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, tips on troubleshooting and avoiding known pitfalls and, where needed, a discussion of the interpretation and use of the results.
Comprehensive and highly practical, E. coli Plasmid Vectors offers those new to the field a basic guide to the use of plasmid vectors in the cloning host E. coli, and those more experienced researchers a broad-ranging, proven array of successful techniques.

Inhaltsverzeichnis

The Function and Organization of Plasmids
Finbarr Hayes

Choosing a Cloning Vector
Andrew Preston

Escherichia coli Host Strains
Nicola Casali

Chemical Transformation of E. coli
W. Edward Swords

Electroporation of E. coli
Claire A. Woodall

DNA Transfer by Bacterial Conjugation
Claire A. Woodall

Cosmid Packaging and Infection of E. coli
Mallory J. A. White and Wade A. Nichols

Isolation of Plasmids from E. coli by Alkaline Lysis
Sabine Ehrt and Dirk Schnappinger

Isolation of Plasmids from E. coli by Boiling Lysis
Sabine Ehrt and Dirk Schnappinger

High-Purity Plasmid Isolation Using Silica Oxide
Stefan Grimm and Frank Voß-Neudecker

High-Throughput Plasmid Extraction Using Microtiter Plates
Michael A. Quail

Isolation of Cosmid and BAC DNA from E. coli
Daniel Sinnett and Alexandre Montpetit

Preparation of Single-Stranded DNA from Phagemid Vectors
W. Edward Swords

Using Desktop Cloning Software to Plan, Track, and Evaluate Cloning Projects
Robert H. Gross

Cloning in Plasmid Vectors
Carey Pashley and Sharon Kendall

Extraction of DNA from Agarose Gels
Nicholas Downey

Cloning PCR Products with T-Vectors
Wade A. Nichols

Construction of Genomic Libraries in l-Vectors
Yilun Wang, Zheng Cao, Darryl Hood, and James G. Townsel

Rapid Screening of Recombinant Plasmids
Sangwei Lu

Restriction Analysis of Recombinant Plasmids
Joanne Goranson-Siekierke and Jarrod L. Erbe

Screening Recombinant DNA Libraries
Wade A. Nichols

Sequencing Using Fluorescent-Labeled Nucleotides
Allison F. Gillaspy

Site-Directed Mutagenesis Using the Megaprimer Method
Zhidong Xu, Alessia Colosimo, and Dieter C. Gruenert

Site-Directed Mutagenesis byInverse PCR
Clifford N. Dominy and David W. Andrews

Creating Nested DNA Deletions Using Exonuclease III
Rosamund Powles and Lafras M. Steyn

Transposon and Transposome Mutagenesis of Plasmids, Cosmids, and BACs
Alistair McGregor

In Vitro Transcription and Translation
Farahnaz Movahedzadeh, Susana González Rico, and Robert A. Cox

Vectors for the Expression of Recombinant Proteins in E. coli
Sally A. Cantrell

Expression of Recombinant Proteins from lac Promoters
Charles R. Sweet

Plasmid-Based Reporter Genes: Assays for b-Galactosidase and Alkaline Phosphatase Activities
Minghsun Liu

Plasmid-Based Reporter Genes: Assays for Green Fluorescent Protein
Sergei R. Doulatov

Index

Pressestimmen

...a scientific review and a methodological "cook-book" of high quality. It is truly a highly practical handbook, useful for all scientists working with gene cloning and manipulation." - Folia Microbiologica
"...a very useful addition to laboratories investigating plasmids per se, and for those employing plasmids as vectors for gene cloning." - Microbiology Today

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