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1. Introduction
2. Physical detection and isolation of plasmid DNA
3. Buoyant density of the F plasmid
4. Topology and ethidium bromide
5. Binding of ethidium bromide to DNA
6. CsCl-ethidium bromide gradients
7. Topology and alkaline denaturation
8. Covalently closed circles renature
9. Plasmid extraction with alkaline lysis
10. ColE1 relaxation complex
11. Electron microscopy of relaxation complex
12. Agarose gel electrophoresis (AGE)
13. Topoisomerases
14. Topoisomers seen on AGE
15. E. coli/ColE1 DNA replication
16. Restriction enzymes
17. Example of restriction enzyme cleavage
18. AGE showing restriction patterns of plasmids
19. AGE patterns reflecting transposon insertions
20. Topoisomers and restriction fragments in AGE
21. Cleavage at BamHI site
22. Ligation of BamHI ends
23. Modification of BamHI site
24. SmaI cleavage
25. Different restriction endonucleases
26. Recombinant DNA technology
27. Cloning DNA fragments with plasmid vector
28. Transformation of plasmid into E. coli
29. Cloning, expression and screening
30. Cloning vectors for E. coli
31. pAM434 with multiple cloning site
32. Shuttle vectors
33. pAM401 shuttle plasmid
34. Sequencing the cloned DNA
35. Map of pAD1
36. Importance of plasmids

Topics Covered

• Properties of plasmids that facilitate isolation
• Basic analyses of plasmids
• Restriction enzymes and their use as analytical tools
• The important role of plasmids in recombinant DNA technology and genome analyses

Links

Series:

• Biology and Significance of Plasmids

Categories:

• Methods
• Microbiology

Therapeutic Areas:

• Infectious Diseases

Talk Citation

Publication History

• Published on January 28, 2008
• Reviewed on April 16, 2023

Financial Disclosures

• Prof. Emeritus Don Clewell has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.