A Laboratory Course

1st Edition - March 28, 1990

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  • Author: Jeffery M. Becker
  • eBook ISBN: 9780323159135

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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

  • Preface


    Suggested Schedule for Exercises

    Introductory Notes

    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

    Exercise 1

    Measurement of pH

    Exercise 2

    Use of Micropipettors and Spectrophotometers

    Exercise 3

    Aseptic Technique: Transferring a Culture

    Exercise 4

    Establishing a Pure Culture: The Streak Plate

    Exercise 5

    Preparation of Culture Media

    Exercise 6

    The Growth Curve

    Exercise 7

    Isolation of Plasmid DNA from Escherichia Coli: The Mini-Prep

    Exercise 8

    Purification, Concentration, and Quantitation of DNA

    Exercise 9

    Isolation of Plasmid DNA: The Maxi-Prep

    Exercise 10

    Restriction Digestion and Agarose Gel Electrophoresis

    Exercise 11

    Southern Transfer

    Exercise 12

    Preparation, Purification, and Hybridization of Probe

    Exercise 13

    Transformation of Saccharomyces cerevisiae

    Exercise 14

    Transformation of Escherichia coli by Plasmid DNA

    Exercise 15

    Protein Assays

    Exercise 16

    β-Galactosidase Assay

    Exercise 17

    Determination of β-Galactosidase in Permeabilized Yeast Cells

    Exercise 18

    Assay of β-Galactosidase in Cell Extracts

    Exercise 19

    β-Galactosidase Purification

    Part A Gel Filtration Chromatography: Column Calibration

    Part Β Ammonium Sulfate Salting-Out of β-Galactosidase and Column Chromatography

    Part C Sodium Dodecyl Sulfate—Polyacrylamide Gel Electrophoresis

    Appendix 1

    Alternative Protocols and Experiments

    Exercise 6A Isolation and Characterization of Auxotrophic Yeast Mutants

    Exercise 9A Large-Scale Isolation of Plasmid DNA by Column Chromatography

    Exercise 12A 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

    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



Product details

  • No. of pages: 250
  • Language: English
  • Copyright: © Academic Press 1990
  • Published: March 28, 1990
  • Imprint: Academic Press
  • eBook ISBN: 9780323159135

About the Author

Jeffery M. Becker

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