Gene Cloning and Manipulation

Part I. The Tools for the Job: 1. Introduction; 2. Cutting; 3. Modification; 4. Ligation; 5. Purification of plasmid DNA from E. Coli; 6. Gel electrophoresis of nucleic acids; 7. Oligonucleotide synthesis; Part II. Simple Cloning: 8. The basic experiment; 9. Vectors; transformation and hosts; 10. Modifications; 11. Linkers, adaptors and cassettes; Part III. Other vector systems for E. Coli; 12. Introduction; 13. Bacteriophage M13 and its uses; 14. Bacteriophage lambda; 15. Cosmids; 16. Bacteriophage P1; Part IV. Making Libraries: 17. Introduction; 18. Genomic libraries; 19. cDNA libraries; 20. Specialized libraries; Part V. Screening Libraries: 21. Introduction; 22. Screening for coding function; 23. Screening for other functions-reporter genes; Part VI. Polymerase Chain Reaction: 24. The basic technique; 25. Modifications; 26. Precautions and drawbacks; 27. Modification and mutagenesis; 28. Introduction; 29. Alteration of restriction sites; 30. Insertions and deletions; 31. Point mutations - early methods; 32. Oligonucleotide mutagenesis; 33. Choosing the right mutations; 34. Inactivating genes; Part VII. Use of Cloned DNA: 35. As DNA; 36. Synthesis of RNA; 37. Synthesis of protein; Part VIII. Using Other Organisms: 38. Gram-negative bacteria; 39. Gram-positive bacteria; 40. Fungi; 41. Chlamydomonas; 42. Vascular plants; 43. Organelle transformation; 44. Insects; 45. Mammals; References and further reading; Index.