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Main description:
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.
Contents:
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 Gonzalez 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
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 316
Weight: 510g
Availability: Available
Subcategories: Biochemistry
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