Biotechnology ：A Laboratory Course ( 2 )

Publication subTitle ：A Laboratory Course

Publication series ：2

Author： Becker   Jeffrey M.;Caldwell   Guy A.;Zachgo   Eve Ann  

Publisher： Elsevier Science‎

Publication year： 1996

E-ISBN: 9780080528199

P-ISBN(Paperback): 9780120845620

P-ISBN(Hardback):  9780120845620

Subject： Q-33 Biological experiments with biological technology

Language： ENG

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Description

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.

  • 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

Chapter

Front Cover

Biotechnology: A Laboratory Course

Copyright Page

Contents

Preface to the Second Edition

Preface to the First Edition

Acknowledgments

Suggested Schedule for Exercises

Introductory Notes: Record Keeping and Safety Rules

Format of Student Laboratory Records

The Ten Commandments of Record Keeping

Safety Rules in the Laboratory

Exercise 1. Aseptic Technique and Establishing Pure Cultures: The Streak Plate and Culture Transfer

Exercise 2. Preparation of Culture Media

Exercise 3. The Growth Curve

Exercise 4. Isolation of Plasmid DNA from Escherichia coli: The Mini-Prep

Exercise 5. Purification, Concentration, and Quantitation of DNA

Exercise 6. Large-Scale Isolation of Plasmid DNA by Column Chromatography

Exercise 7. Amplification of a lacZ Gene Fragment by the Polymerase Chain Reaction

Exercise 8. Restriction Digestion and Agarose Gel Electrophoresis

Exercise 9. Southern Transfer

Exercise 10. Preparation, Purification, and Hybridization of Probe

Exercise 11. Transformation of Saccharomyces cerevisiae

Exercise 12. Isolation of Plasmid from Yeast and Escherichia coil Transformation

Exercise 13. Protein Assays

Exercise 14. Qualitative Assay for b-Galactosidase in Yeast Colonies

Exercise 15. Determination of b-Galactosidase in Permeabilized Yeast Cells

Exercise 16. Assay of b-Galactosidase in Cell Extracts

Exercise 17. b-Galactosidase Purification

Exercise 18. Western Blot: Probe of Protein Blot with Antibody to b-Galactosidase

Appendix 1. Alternative Protocols and Experiments

Exercise 1A Isolation and Characterization of Auxotrophic Yeast Mutants

Exercise 2A Measurement of pH

Exercise 3A Use of the Spectrophotometer

Exercise 6A Isolation of Plasmid DNA: The Maxi-Prep

Exercise 10A Colony Hybridization

Appendix 2. Buffer Solutions

Appendix 3. Preparation of Buffers and Solutions

Appendix 4. Properties of Some Common Concentrated Acids and Bases

Appendix 5. Use of Micropipettors

Appendix 6. Safe Handling of Microorganisms

Appendix 7. List of Cultures

Appendix 8. Storage of Cultures and DNA

Appendix 9. Sterilization Methods

Appendix 10. Preparation of Stock Solutions for Culture Media

Appendix 11. Growth in Liquid Medium

Appendix 12. Determination of Viable Cells

Appendix 13. Determination of Cell Mass

Appendix 14. Determination of Cell Number

Appendix 15. Nomenclature of Strains

Appendix 16. Glassware and Plasticware

Appendix 17. Preparation of Tris and EDTA

Appendix 18. Basic Rules for Handling Enzymes

Appendix 19. Effects of Common Contaminants on Protein Assays

Appendix 20. Manufacturers' and Distributors' Addresses

Appendix 21. Surfing the Bionet: World Wide Web Addresses

Glossary

Index

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