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List of contents
Introduction; Acknowledgements; Laboratory Rules and Safety; Experiment 1: An exercise in pipetting and the Beer-Lambert Law; Experiment 2: Bacterial growth curve; Experiment 3: Bioassay for micotinic acid; Experiment 4: The effect of environment on growth; Experiment 5: Lactic acid production by lactic acid bacteria; Experiment 6: Assay of protein and RNA in whole cells grown at different growth rates; Experiment 7: Analysis of diauxic growth; Experiment 8: Assay of amylase and protease secreted by Bacillus Subtilis; Experiment 9: Concentration of amylase from Bacillus Subtilis by ammonium sulfate precipitation and separation from protease by affinity purification; Experiment 10: Ion-exchange chromatography of amylase; Experiment 11: Induction of alkaline phosphatase and the determination of its cellular cation in Escherichia coli; Experiment 12: Assay of threonine deaminase: Determination of KM and Vmax; Experiment 13: Assaying fructose-1,6-bisphosphatase in Saccharomyces cerevisiae; Experiment 14: Purification of glucose-6-phosphate dehydrogenase; Experiment 15: How E. coli adapts to anaerobiosis: Nitrate reductase; Experiment 17: Chemotaxis of Pseudomonas aeruginosa; Experiment 18: Phototasix of Rhodospirllum centenum; Experiment 19: Light production by Phorobacterium phosphoreum; Appendix A: Analysis of experimental data: How to do simple dilutions; Guidelines for writing laboratory reports; Appendix B: Laboratory supplies and equipment; Appendix C: Media; Buffers; Solutions; Appendix D: RNA assay using orcinol; Lowry protein assay; Bradford Protein assay; Assay for *a-amylase using starch conjugated to a dye; Appendix E: The Beer-Lambert law; Relative absorbancies; Cell density vs. Klett units; Appendix F: Quantitative problems; Appendix G: Solutions to problems in Appendix F; Appendix H: Determining the amount of inorganic phosphate that limits the growth yields of E. coli, Factors that affect the relative amounts of saturated; unsaturated, and branched chain fatty acids; Measuring the KM and Vmax of fructose-1,6-bisphosphatase and the inhibition by AMP; Assaying glycolytic enzymes and fructose-1,6-bisphosphatase in yeast grown on glucose or ethanol; Regulation of phosphofructokinase activity; Further experiments and demonstrations with Photobacterium
Summary
This laboratory manual demonstrating microbial diversity is intended as an adjunct to the author's "Physiology and Biochemistry of Prokaryotes", but can also be used in conjunction with other books. It includes 18 experiments for students to conduct in the laboratory.
