Biotechnology

Preface

Acknowledgments

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

Glossary

Index