E. coli plasmid vectors : methods and applications /

Bibliographic Details
Other Authors: Casali, Nicola, Preston, Andrew, 1969-
Format: Book
Language:English
Published: Totowa, N.J. : Humana Press, [2003]
Series:Methods in molecular biology (Clifton, N.J.) ; v. 235.
Subjects:
Table of Contents:
  • Function and organization of plasmids
  • Choosing a cloning vector
  • Escherichia coli host strains
  • Chemical transformation of E. coli
  • Electroporation of E. coli
  • DNA transfer by bacterial conjugation
  • Cosmid packaging and infection of E. coli
  • Isolation of plasmids from E. coli by boiling lysis
  • High-purity plasmid isolation using silica oxide
  • High-throughput plasmid extraction using microtiter plates
  • Isolation of cosmid and BAC DNA from E. coli
  • Preparation of single-stranded DNA from phagemid vectors
  • Using desktop cloning software to plan, track, and evaluate cloning projects
  • Cloning in plasmid vectors
  • Extraction of DNA from agarose gels
  • Cloning PCR products with T-vectors
  • Rapid screening of recombinant plasmids
  • Restriction analysis of recombinant plasmids
  • Screening recombinant DNA libraries
  • Sequencing using fluorescent-labeled nucleotides
  • Site-directed mutagenesis using the megaprimer method
  • Site-directed mutagenesis by inverse PCR
  • Creating nested DNA deletions using exonuclease III
  • Transposon and transposome mutagenesis of plasmids, cosmids, and BACs
  • In vitro transcription and translation
  • Vectors for the expression of recombinant proteins in E. coli
  • Expression of recombinant proteins form Iac promoters
  • Plasmid-based reporter genes : assays for green fluorescent protein