MCB 4403L PROKARYOTIC BIOLOGY LABORATORY Summer 2007 Laboratory Syllabus

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1 MCB 4403L PROKARYOTIC BIOLOGY LABORATORY Summer 2007 Laboratory Syllabus Week 1 1 5/09 (W) - Orientation: Laboratory Operation and Procedures, Grading and Attendance Policies, 5/10 (R) and Laboratory Safety Rules TOPICS: Handling Microorganisms Safely and Sterilization Techniques Begin: Ex. 1 = Principles of Aseptic Technique (p. 3) Ex. 2 = Aseptic Method for Entering Culture Tubes and Transferring Cells (p. 5) Ex. 12 = Sterilization Principles and Methods (p. 99) Week 2 2 5/14 (M) - TOPICS: Cell Morphology, Staining Microorganisms, and Introduction to Microscopy 5/15 (T) Begin: Ex. 3 = Principles and Care of the Light Microscope (p. 13) Ex. 4 = Introduction to Staining Microorganisms (p. 21) Ex. 5 = Smear Preparation, Fixation, and Simple Staining with Basic Dyes (p. 27) Ex. 13 = Preparing Culture Media (p. 107) 3 5/16 (W) - TOPICS: Differential Staining 5/17 (R) Begin: Ex. 7A = The Gram Stain: A Differential Stain (p. 51) Ex. 13 = Preparing Culture Media (p. 107) Week 3 4 5/21(M) - TOPICS: Motility and Structural Staining 5/22(T) Begin: Ex. 9A = Bacterial Endospores: Observing Endospores with a Phase-Contrast Microscope (p. 69) - Demonstration Ex. 9B = Bacterial Endospores: Observing Endospores with Simple Stains and The Gram Stain (p. 69) Ex. 9C = Bacterial Endospores: Structural Staining of Endospores with Hot Malachite Green (p. 70) Ex.10B= Bacterial Flagella and Motility: Observing Motility with the Phase-Contrast Microscope - Demonstration 5 5/23 (W) - TOPICS: Culturing and Enumerating Bacteria: Serial Dilution and Viable Counts 5/24(R) Begin: Ex. 14 = Separating Microbes on Streak Plates (p. 119) Ex. 16 = Agar-Slant and Agar-Deep Cultures (p. 143) Ex. 17 = Broth Cultures (p. 151) (**24 hr. observations) Ex. 20 = Counting Viable Cells: Serial Dilution and Spread Plates (p.175) **IMPORTANT NOTE: Ex. 17 requires that some of the results be recorded after 24 hours. This means that all students will have to come into the lab (for a brief period of time) on the day after they start Ex. 17 (i.e., on a day they normally do not have lab), in order to take this 24-hour reading. Additional information will be provided by the instructors during the regular lab periods.

2 MCB4403L, Syllabus (cont.) Week 4 5/28 (M)- No Classes Memorial Day Holiday 5/29 (T) 6 5/30 (W)- TOPICS: Determining Cultural Purity 5/31 (R)- Begin: Ex. 7B = Gram stain of an unknown microorganism (p.54) Ex. 15A= Determining Culture Purity: Testing the Purity of a Colony (p.131) Ex. 14 = Separating Microbes on Streak Plates (p. 119) Ex. 16 = Agar-Slant and Agar-Deep Cultures (p. 143) Ex. 17 = Broth Cultures (p. 151) Ex. 20 = Counting Viable Cells: Serial Dilution and Spread Plates (p.175) Week 5 7 6/04 (M)- TOPICS: Selective Culturing and QUIZ #1 6/05 (T) Begin: Ex. 21 = Selective and Differential Media (p. 195) Ex. 15A= Determining Culture Purity: Testing the Purity of a Colony (p. 131) 8 6/06(W)- TOPICS: Environmental Effects on Growth and 6/07 (R) LABORATORY REPORT #1 DUE: GRAM STAIN Begin: Ex. 18 = Culturing Anaerobes: The Anaerobe Jar (p. 159) Ex. 23 = The Effects of Temperature on Growth (p. 213) Ex. 24 = The Effects of Elevated Sugar and NaCl Concentrations on Growth (p. 219) Ex. 25 = The Effects of Free-Oxygen Concentration on Growth: Agar-Deep Cultures (p. 225) Week 6 9 6/11 (M)- TOPICS: Viruses and Molecular Biology 6/12 (T) Begin: Ex. 37 = Enumeration of Lytic Viruses: The Plaque Assay (p.315) Ex. 39A = Effects of Ultraviolet Light on DNA and Cell Viability (p. 333) Ex. 18 = Culturing Anaerobes: The Anaerobe Jar (p.159) Ex. 23 = The Effects of Temperature on Growth (p. 213) Ex. 24 = The Effects of Elevated Sugar and Sodium Chloride Concentrations on Growth (p. 219) Ex. 25 = The Effects of Free-Oxygen Concentration on Growth: Agar-Deep Cultures (p. 225) 10 6/13 (W) - TOPICS: Nutrient Degradation and Transport 6/14 (R) Begin: Ex. 26 = Introduction to Extracellular Degradation (p. 235) Ex. 27 = Microbial Degradation of Polysaccharides Starch (p.241) Ex. 28 = Microbial Degradation of Proteins Casein and Gelatin (p. 247) Ex. 29 = Microbial Degradation of Lipids (p. 257) Ex. 37 = Enumeration of Lytic Viruses: The Plaque Assay Ex. 39A = Effects of Ultraviolet Light on DNA and Cell Viability (p. 333)

3 Week /18 (M) - TOPICS: Metabolic Products and QUIZ #2 6/19 (T) Begin: Ex. 30 = Differential Utilization of Citrate by the Enteric Bacteria (p. 263) Ex. 31 = Acid and Gas Production from Sugar Fermentation (p. 269) Ex. 32 = Acid and Neutral Products from Sugar Fermentation: The Methyl Red and Voges-Proskauer Tests (p. 277) Ex. 34 = Indole Production from the Amino Acid Tryptophan and Catabolite Repression (p. 291) Ex. 27 = Microbial Degradation of Polysaccharides Starch (p. 241) Ex. 28 = Microbial Degradation of Proteins Casein and Gelatin (p. 241) Ex. 29 = Microbial Degradation of Lipids (p. 257) 12 6/20 (W) - TOPICS: Metabolic Products and LABORATORY REPORT #2 DUE: TBA 6/21 (R) Begin: Ex. 33 = Microbial Hydrogen Sulfide (H2S) Production from Thiosulfate and Sulfur-Containing Amino Acids (p. 285) Ex. 35 = Products Formed in Milk: The Litmus Milk Test (p.297) (**24 hr observations) Ex. 36 = Test for Cytochrome c (Oxidase) and Catalase Activities (p.303) Ex. 28 = Microbial Degradation of Proteins Casein and Gelatin Ex. 30 = Differential Utilization of Citrate by the Enteric Bacteria Ex. 31 = Acid and Gas Production from Sugar Fermentation Ex. 32 = Acid and Neutral Products from Sugar Fermentation: The Methyl Red and Voges-Proskauer Tests (p. 277) Ex. 34 = Indole Production from the Amino Acid Tryptophan and Catabolite Repression Week /25 (M) - TOPICS: Environmental Microbiology and Food Microbiology 6/26(T) Begin: Ex. 50 = Milk Preservation: Yogurt (p. 455) Ex. 52 = Numbers of Bacteria on Solid Foods (p. 469) Ex. 33 = Microbial Hydrogen Sulfide (H2S) Production from Thiosulfate and Sulfur-Containing Amino Acids (p. 285) Ex. 35 = Products Formed in Milk: The Litmus Milk Test (p.297) **IMPORTANT NOTE: Ex. 35 requires that some of the results be recorded after 24 hours. This means that all students will have to come into the lab (for a brief period of time) on the day after they start Ex. 35 (i.e., on a day they normally do not have lab), in order to take this 24-hour reading. Additional information will be provided by the instructors during the regular lab periods.

4 14 6/27 (W) - TOPICS: Medical Microbiology and Immunology 6/28 (R) Begin: *SE#1 =Transmission of Microbes on Human Skin and Fomites (after appendices) Ex. 8 =The Acid-Fast Stain: A Differential Stain Ex.15B = Determining Culture Purity: Separating Cultures from an Unknown Mixture(p.131) Ex. 45 = Coagulase Production by Pathogenic Staphylococci (p.413) Ex. 46 = Hemolysis of Red Blood Cells (p. 421) Ex. 35 = Products Formed in Milk: The Litmus Milk Test (p.297) Ex. 50 = Milk Preservation: Yogurt (p.455) Ex. 52 = Numbers of Bacteria on Solid Foods (p. 469) Week /02 (M) - TOPICS: Medical Microbiology and Immunology and QUIZ #3 7/03(T) Begin: Ex. 47 = Bacteria on Human Skin (p. 431) Ex. 48 = Bacteria in the Human Throat (p. 441) *SE#2 = Bacteria in Human Urine (after appendices) *SE #1 = Transmission of Microbes on Human Skin and Fomites (after appendices) Ex.15B = Determining Culture Purity: Testing the Purity of a Colony Ex. 46 = Hemolysis of Red Blood Cells (p. 421) 7/04 (W)- No Classes Fourth of July Holiday 7/05 (R)- Week /09 (M) TOPICS: Medical Microbiology and Immunology LABORATORY REPORT #3 DUE: TBA 7/10 (T) Begin: Ex. 43 = Evaluating the Effectiveness of Common Antiseptics and Disinfectants (p. 397) Ex. 44 = Antibiotic Evaluation by the Kirby-Bauer Method (p.405) Ex. 58 = Identification of Unknown Microorganisms (p. 515) Ex. 47 = Bacteria on Human Skin (p. 431) Ex. 48 = Bacteria in the Human Throat (p. 441) *SE#2 = Bacteria in Human Urine (after appendices) 17 7/11 (W) TOPICS: Molecular Techniques 7/12 (R) Begin: *SE #3 = DNA Fingerprinting and Phylogenetic Analysis of Bacteria (after appendices) Ex. 43 = Evaluating the Effectiveness of Common Antiseptics and Disinfectants (p. 397) Ex. 44 = Antibiotic Evaluation by the Kirby-Bauer Method (p.405) *SE #2 = Bacteria in Human Urine (after appendices) Ex. 58 = Identification of Unknown Microorganisms (p. 515)

5 Week /16(M) QUIZ #4 7/17 (T) Ex. 51 = Solid Food Preservation Sauerkraut (p. 463) Ex. 58 = Identification of Unknown Microorganisms (p.515) *SE #3 = DNA Fingerprinting and Phylogenetic Analysis of Bacteria (see website) 19 7/18 (W) - TOPICS: Bacterial Genetics LABORATORY REPORT #4 DUE TBA 7/19(R) Begin: *SE #4 = Transformation of Bacteria (see website) Ex. 58 = Identification of Unknown Microorganisms (p.515) *SE #3 = DNA Fingerprinting and Phylogenetic Analysis of Bacteria (see website) Week /23 (M) 7/24 (T) Ex. 58 = Identification of Unknown Microorganisms (p. 515) *SE #4 = Transformation of Bacteria (see website) 7/25(W) 7/26(R) QUIZ #5 AND FINAL EXAMINATION (LAB PRACTICUM EXAM) Lab notebooks due at end of class.period. 7/27 (F) Written report on identification of unknown due by 5:00pm This document and all others produced for this course are available upon request in alternate formats for individuals with print-related disabilities. Contact Dr. Reeves at for more information. *Supplemental Exercises will be posted on the FSU Biological Science Page under Course Materials and then MCB4403L (http://www.bio.fsu.edu/index-course_materials.php). Students will have to download and print these exercises prior to the date these exercises will be performed.

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