Table of Contents

The 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 Alkaline Lysis.- 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.- Construction of Genomic Libraries in ?-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 From lac Promoters.- Plasmid-Based Reporter Genes.- Plasmid-Based Reporter Genes.

Reviews

...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