Biotechnology

A Laboratory Course

Jeffery M. Becker

• Publisher: Academic Press
• Year: 2012
• Print ISBN: 9780120845606
• eBook ISBN: 9780323159135
• Topic: Tecnologia e ingegneria
• Subtopic: Chimica industriale e tecnica

Biotechnology: A Laboratory Course is a series of laboratory exercises demonstrating the in-depth experience and understanding of selected methods, techniques, and instrumentation used in biotechnology. This manual is an outgrowth of an introductory laboratory course for senior undergraduate and first year graduate students in the biological sciences at The University of Tennessee. This book is composed of 19 chapters and begins with some introductory notes on record keeping and safety rules. The first exercises include pH measurement, the use of micropipettors and spectrophotometers, the concept of aseptic technique, and preparation of culture media. The subsequent exercises involve the application of the growth curve, the isolation, purification, and concentration of plasmid DNA from Escherichia coli, and the process of agarose gel electrophoresis. Other exercises include the preparation, purification, and hybridization of probe, the transformation of Saccharomyces cerevisiae, the transformation of E. coli by plasmid DNA, and the principles and applications of protein assays. The final exercises explore the?-galactosidase assay and the purification and determination of?-galactosidase in permeabilized yeast cells. This book is of great value to undergraduate biotechnology and molecular biology students.

Table of contents

• Front Cover
• Biotechnology: A Laboratory Course
• Copyright Page
• Table of Contents
• Dedication
• Preface
• Acknowledgments
• Suggested Schedule for Exercises
• Record Keeping and Safety Rules
• Format of Student Laboratory Records
• Ten Dos and Don'ts of Record Keeping
• Criteria for Grading the Laboratory Notebook
• Safety Rules in the Laboratory
• Chapter 1. Measurement of pH
• Chapter 2. Use of Micropipettors and Spectrophotometers
• Chapter 3. Aseptic Technique: Transferring a Culture
• Chapter 4. Establishing a Pure Culture: The Streak Plate
• Chapter 5. Preparation of Culture Media
• Chapter 6. The Growth Curve
• Chapter 7. Isolation of Plasmid DNA from Escherichia coli: The Mini-Prep
• Chapter 8. Purification, Concentration, and Quantitation of DNA
• Chapter 9. Isolation of Plasmid DNA: The Maxi-Prep
• Chapter 10. Restriction Digestion and Agarose Gel Electrophoresis
• Chapter 11. Southern Transfer
• Chapter 12. Preparation, Purification, and Hybridization of Probe
• Chapter 13. Transformation of Saccharomyces cerevisiae
• Chapter 14. Transformation of Escherichia coli by Plasmid DNA
• Chapter 15. Protein Assays
• Chapter 16. β-Galaetosidase Assay
• Chapter 17. Determination of β-Galactosidase in Permeabilized Yeast Cells
• Chapter 18. Assay of β-Galactosidase in Cell Extracts
• Chapter 19. β-Galactosidase Purification
• Appendix 1. Alternative Protocolsand Experiments
• 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
• 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. Manufacturers' and Distributors' Addresses
• Glossary
• Index