E. coli Plasmid Vectors

1. Front Matter

   Pages i-xi

   PDF

2. The Function and Organization of Plasmids

   • Finbarr Hayes

   Pages 1-17

3. Choosing a Cloning Vector

   • Andrew Preston

   Pages 19-26

4. Escherichia coli Host Strains

   • Nicola Casali

   Pages 27-48

5. Chemical Transformation of E. coli

   • W. Edward Swords

   Pages 49-53

6. Electroporation of E. coli

   • Claire A. Woodall

   Pages 55-59

7. DNA Transfer by Bacterial Conjugation

   • Claire A. Woodall

   Pages 61-65

8. Cosmid Packaging and Infection of E. coli

   • Mallory J. A. White, Wade A. Nichols

   Pages 67-73

9. Isolation of Plasmids from E. coli by Alkaline Lysis

   • Sabine Ehrt, Dirk Schnappinger

   Pages 75-78

10. Isolation of Plasmids from E. coli by Boiling Lysis

    • Sabine Ehrt, Dirk Schnappinger

    Pages 79-82

11. High-Purity Plasmid Isolation Using Silica Oxide

    • Stefan Grimm, Frank Voß-Neudecker

    Pages 83-87

12. High-Throughput Plasmid Extraction Using Microtiter Plates

    • Michael A. Quail

    Pages 89-97

13. Isolation of Cosmid and BAC DNA from E. coli

    • Daniel Sinnett, Alexandre Montpetit

    Pages 99-102

14. Preparation of Single-Stranded DNA from Phagemid Vectors

    • W. Edward Swords

    Pages 103-106

15. Using Desktop Cloning Software to Plan, Track, and Evaluate Cloning Projects

    • Robert H. Gross

    Pages 107-120

16. Cloning in Plasmid Vectors

    • Carey Pashley, Sharon Kendall

    Pages 121-135

17. Extraction of DNA from Agarose Gels

    • Nicholas Downey

    Pages 137-139

18. Cloning PCR Products with T-Vectors

    • Wade A. Nichols

    Pages 141-152

19. Construction of Genomic Libraries in λ-Vectors

    • Yilun Wang, Zheng Cao, Darryl Hood, James G. Townsel

    Pages 153-168

20. Rapid Screening of Recombinant Plasmids

    • Sangwei Lu

    Pages 169-174

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.

• BAC
• DNA
• Escherichia coli
• PCR
• Promoter
• Translation
• chromosome
• genes
• reproducible techniques
• transcription

...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

• University of California at Berkeley, School of Public Health, Berkeley

  Nicola Casali

• Department of Clinical Veterinary Medicine, University of Cambridge, Cambridge, UK

  Andrew Preston

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