Elsevier S & T

Biotechnology: A Laboratory Course

Becker, Jeffrey M.; Caldwell, Guy A.; Zachgo, Eve Ann

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Table of contents
  • Contentsvii
  • Preface to the Second Editionxiii
  • Preface to the First Editionxv
  • Acknowledgmentsxvii
  • Suggested Schedule for Exercisesxix
  • Introductory Notes: Record Keeping and Safety Rules1
  • Format of Student Laboratory Records3
  • The Ten Commandments of Record Keeping5
  • Safety Rules in the Laboratory7
  • Exercise 1. Aseptic Technique and Establishing Pure Cultures: The Streak Plate and Culture Transfer9
  • Exercise 2. Preparation of Culture Media17
  • Exercise 3. The Growth Curve25
  • Exercise 4. Isolation of Plasmid DNA from Escherichia coli: The Mini-Prep31
  • Exercise 5. Purification, Concentration, and Quantitation of DNA37
  • Exercise 6. Large-Scale Isolation of Plasmid DNA by Column Chromatography49
  • Exercise 7. Amplification of a lacZ Gene Fragment by the Polymerase Chain Reaction55
  • Exercise 8. Restriction Digestion and Agarose Gel Electrophoresis63
  • Exercise 9. Southern Transfer75
  • Exercise 10. Preparation, Purification, and Hybridization of Probe85
  • Exercise 11. Transformation of Saccharomyces cerevisiae97
  • Exercise 12. Isolation of Plasmid from Yeast and Escherichia coil Transformation105
  • Exercise 13. Protein Assays119
  • Exercise 14. Qualitative Assay for β -Galactosidase in Yeast Colonies125
  • Exercise 15. Determination of b-Galactosidase in Permeabilized Yeast Cells131
  • Exercise 16. Assay of β -Galactosidase in Cell Extracts135
  • Exercise 17. β -Galactosidase Purification141
  • Exercise 18. Western Blot: Probe of Protein Blot with Antibody to β -Galactosidase157
  • Appendix 1. Alternative Protocols and Experiments163
  • Exercise 1A Isolation and Characterization of Auxotrophic Yeast Mutants165
  • Exercise 2A Measurement of pH170
  • Exercise 3A Use of the Spectrophotometer174
  • Exercise 6A Isolation of Plasmid DNA: The Maxi-Prep178
  • Exercise 10A Colony Hybridization189
  • Appendix 2. Buffer Solutions192
  • Appendix 3. Preparation of Buffers and Solutions195
  • Appendix 4. Properties of Some Common Concentrated Acids and Bases200
  • Appendix 5. Use of Micropipettors201
  • Appendix 6. Safe Handling of Microorganisms204
  • Appendix 7. List of Cultures207
  • Appendix 8. Storage of Cultures and DNA208
  • Appendix 9. Sterilization Methods210
  • Appendix 10. Preparation of Stock Solutions for Culture Media212
  • Appendix 11. Growth in Liquid Medium214
  • Appendix 12. Determination of Viable Cells217
  • Appendix 13. Determination of Cell Mass219
  • Appendix 14. Determination of Cell Number221
  • Appendix 15. Nomenclature of Strains223
  • Appendix 16. Glassware and Plasticware226
  • Appendix 17. Preparation of Tris and EDTA228
  • Appendix 18. Basic Rules for Handling Enzymes231
  • Appendix 19. Effects of Common Contaminants on Protein Assays235
  • Appendix 20. Manufacturers' and Distributors' Addresses237
  • Appendix 21. Surfing the Bionet: World Wide Web Addresses245
  • Glossary247
  • Index253
Book details
  • Vendor Elsevier S & T
  • SKU 9780120845620
  • ISBN-13 9780080528199
  • Author Becker, Jeffrey M.; Caldwell, Guy A.; Zachgo, Eve Ann
  • Edition 2nd
  • Category Science
  • Subject Biotechnology

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Description eBook delivery & access Reviews
The objectives of this Second Edition of Biotechnology: A Laboratory Course remain unchanged: to create a text that consists of a series of laboratory exercises that integrate molecular biology with protein biochemistry techniques while providing a continuum of experiments. The course begins with basic techniques and culminates in the utilization of previously acquired technical experience and experimental material. Two organisms, Sacchaomyces cerevisiae and Escherichia coli, a single plasmid, and a single enzyme are the experimental material, yet the procedures and principles demonstrated are widely applicable to other systems. This text will serve as an excellent aid in the establishment or instruction of introductory courses in the biological sciences.

Key Features of this new edition:
* All exercises and appendixes have been updated
* Includes new exercises on
* Polymerase chain reaction
* Beta-Galactosidase detection in yeast colonies
* Western blotting
* New procedures introduced for
* Large-scale plasmid isolation
* Yeast transformation
* DNA quantitation
* New appendixes added, one of which provides details on accessing biological information sites on the Internet (World Wide Web)
* Use of non-radioactive materials and easy access to microbial cultures
* Laboratory exercises student tested for seven years