Biotechnology A Laboratory Course [2 ed.] 0120845628, 9780120845620, 9780080528199

Table of contents :
Front Cover......Page 1
Biotechnology: A Laboratory Course......Page 4
Copyright Page......Page 5
Contents......Page 8
Preface to the Second Edition......Page 14
Preface to the First Edition......Page 16
Acknowledgments......Page 18
Suggested Schedule for Exercises......Page 20
Introductory Notes: Record Keeping and Safety Rules......Page 22
Format of Student Laboratory Records......Page 24
The Ten Commandments of Record Keeping......Page 26
Safety Rules in the Laboratory......Page 28
Exercise 1. Aseptic Technique and Establishing Pure Cultures: The Streak Plate and Culture Transfer......Page 30
Exercise 2. Preparation of Culture Media......Page 38
Exercise 3. The Growth Curve......Page 46
Exercise 4. Isolation of Plasmid DNA from Escherichia coli: The Mini-Prep......Page 52
Exercise 5. Purification, Concentration, and Quantitation of DNA......Page 58
Exercise 6. Large-Scale Isolation of Plasmid DNA by Column Chromatography......Page 70
Exercise 7. Amplification of a lacZ Gene Fragment by the Polymerase Chain Reaction......Page 76
Exercise 8. Restriction Digestion and Agarose Gel Electrophoresis......Page 84
Exercise 9. Southern Transfer......Page 96
Exercise 10. Preparation, Purification, and Hybridization of Probe......Page 106
Exercise 11. Transformation of Saccharomyces cerevisiae......Page 118
Exercise 12. Isolation of Plasmid from Yeast and Escherichia coil Transformation......Page 126
Exercise 13. Protein Assays......Page 140
Exercise 14. Qualitative Assay for b-Galactosidase in Yeast Colonies......Page 146
Exercise 15. Determination of b-Galactosidase in Permeabilized Yeast Cells......Page 152
Exercise 16. Assay of b-Galactosidase in Cell Extracts......Page 156
Exercise 17. b-Galactosidase Purification......Page 162
Exercise 18. Western Blot: Probe of Protein Blot with Antibody to b-Galactosidase......Page 178
Appendix 1. Alternative Protocols and Experiments......Page 184
Exercise 1A Isolation and Characterization of Auxotrophic Yeast Mutants......Page 186
Exercise 2A Measurement of pH......Page 191
Exercise 3A Use of the Spectrophotometer......Page 195
Exercise 6A Isolation of Plasmid DNA: The Maxi-Prep......Page 199
Exercise 10A Colony Hybridization......Page 210
Appendix 2. Buffer Solutions......Page 213
Appendix 3. Preparation of Buffers and Solutions......Page 216
Appendix 4. Properties of Some Common Concentrated Acids and Bases......Page 221
Appendix 5. Use of Micropipettors......Page 222
Appendix 6. Safe Handling of Microorganisms......Page 225
Appendix 7. List of Cultures......Page 228
Appendix 8. Storage of Cultures and DNA......Page 229
Appendix 9. Sterilization Methods......Page 231
Appendix 10. Preparation of Stock Solutions for Culture Media......Page 233
Appendix 11. Growth in Liquid Medium......Page 235
Appendix 12. Determination of Viable Cells......Page 238
Appendix 13. Determination of Cell Mass......Page 240
Appendix 14. Determination of Cell Number......Page 242
Appendix 15. Nomenclature of Strains......Page 244
Appendix 16. Glassware and Plasticware......Page 247
Appendix 17. Preparation of Tris and EDTA......Page 249
Appendix 18. Basic Rules for Handling Enzymes......Page 252
Appendix 19. Effects of Common Contaminants on Protein Assays......Page 256
Appendix 20. Manufacturers' and Distributors' Addresses......Page 258
Appendix 21. Surfing the Bionet: World Wide Web Addresses......Page 266
Glossary......Page 268
Index......Page 274