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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.
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
Measurement of pH
Use of Micropipettors and Spectrophotometers
Aseptic Technique: Transferring a Culture
Establishing a Pure Culture: The Streak Plate
Preparation of Culture Media
The Growth Curve
Isolation of Plasmid DNA from Escherichia Coli: The Mini-Prep
Purification, Concentration, and Quantitation of DNA
Isolation of Plasmid DNA: The Maxi-Prep
Restriction Digestion and Agarose Gel Electrophoresis
Preparation, Purification, and Hybridization of Probe
Transformation of Saccharomyces cerevisiae
Transformation of Escherichia coli by Plasmid DNA
Determination of β-Galactosidase in Permeabilized Yeast Cells
Assay of β-Galactosidase in Cell Extracts
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
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
Preparation of Buffers and Solutions
Properties of Some Common Concentrated Acids and Bases
Use of Micropipettors
Safe Handling of Microorganisms
List of Cultures
Storage of Cultures
Preparation of Stock Solutions for Culture Media
Growth in Liquid Medium
Determination of Viable Cells
Determination of Cell Mass
Determination of Cell Number
Nomenclature of Strains
Glassware and Plasticware
Preparation of Tris and EDTA
Basic Rules for Handling Enzymes
Manufacturers' and Distributors' Addresses
- No. of pages:
- © Academic Press 1990
- 28th March 1990
- Academic Press
- eBook ISBN:
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