Biotechnology

Suggested Schedule for Laboratory 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 Culture and Establishing Pure Cultures.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 coli Transformation.Exercise 13: Protein Assays.Exercise 14: Qualitative Assay for Galactosidase in Yeast Colonies.Exercise 15: Determination of Galactosidase in Permeabilized Yeast Cells.Exercise 16: Assay of Galactosidase in Cell Extracts.Exercise 17: Galactosidase Purification.Exercise 18: Western Blot: Probe of Protein Blot with Antibody to Galactosidase.Appendix 1: Alternate 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.Subject Index.