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Suggested Schedule for Laboratory Exercises | |
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Introductory Notes | |
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Record Keeping and Safety Rules | |
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Format of Student Laboratory Records | |
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The Ten Commandments of Record Keeping | |
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Safety Rules in the Laboratory | |
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Exercise 1: Aseptic Culture and Establishing Pure Cultures | |
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Exercise 2: Preparation of Culture Media | |
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Exercise 3: The Growth Curve | |
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Exercise 4: Isolation of Plasmid DNA from Escherichia coli: The Mini-Prep | |
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Exercise 5: Purification, Concentration, and Quantitation of DNA. Exercise 6: Large-Scale Isolation of Plasmid DNA by Column Chromatography | |
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Exercise 7: Amplification of a lacZ | |
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Gene Fragment by the Polymerase Chain Reaction | |
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Exercise 8: Restriction Digestion and Agarose Gel Electrophoresis | |
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Exercise 9: Southern Transfer | |
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Exercise 10: Preparation, Purification, and Hybridization of Probe | |
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Exercise 11: Transformation of Saccharomyces cerevisiae | |
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Exercise 12: Isolation of Plasmid from Yeast and Escherichia coli Transformation | |
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Exercise 13: Protein Assays | |
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Exercise 14: Qualitative Assay for Galactosidase in Yeast Colonies | |
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Exercise 15: Determination of Galactosidase in Permeabilized Yeast Cells | |
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Exercise 16: Assay of Galactosidase in Cell Extracts | |
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Exercise 17: Galactosidase Purification | |
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Exercise 18: Western Blot: Probe of Protein Blot with Antibody to Galactosidase | |
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Appendix 1: Alternate Protocols and Experiments: Exercise 1A: Isolation and Characterization of Auxotrophic Yeast Mutants | |
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Exercise 2A: Measurement of pH. Exercise 3A: Use of the Spectrophotometer | |
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Exercise 6A: Isolation of Plasmid DNA: The Maxi-Prep | |
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Exercise 10A: Colony Hybridization | |
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: Buffer Solutions | |
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Preparation of Buffers and Solutions | |
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Properties of Some Common Concentrated Acids and Bases | |
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Use of Micropipettors | |
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Safe Handling of Microorganisms | |
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List of Cultures | |
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Storage of Cultures and DNA | |
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Sterilization Methods | |
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Preparation of Stock Solutions for Culture Media | |
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Growth in Liquid Medium | |
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Determination of Viable Cells | |
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Determination of Cell Mass | |
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Determination of Cell Number | |
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Nomenclature of Strains | |
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Glassware and Plasticware | |
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Preparation of Tris and EDTA | |
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Basic Rules for Handling Enzymes | |
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Effects of Common Contaminants on Protein Assays | |
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Manufacturers' and Distributors' Addresses | |
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Surfing the Bionet: World Wide Web Addresses | |
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Glossary | |
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Subject Index | |