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Bio110AntibioticResistanceLabManual2013_

Bio110AntibioticResistanceLabManual2013_ - (s)ofBacterial...

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1 Bio 110 Laboratory Activity Using Genes for Antibiotic Resistance to Trace Source(s) of Bacterial Contamination Week 1: The Serial Dilution Method for Counting Populations of Bacteria to Week 2: Using PCR to Identify Antibiotic Resistance Genes Goals: 1. Understand that bacterial antibiotic resistance genes are often carried on plasmids. 2. Understand that antibiotic resistance genes spread quickly through bacterial populations due to the nature of the plasmids themselves, how they replicate, and how they are transferred to other bacteria. 3. Become familiar with the use of antibiotics to screen for antibiotic resistance. 4. Understand that kanamycin operates by interfering with protein synthesis at a site on the bacterial ribosome. 5. Understand the theory behind using serial dilution to determine the concentration of bacteria in a suspension. 6. Understand why determining the frequency of kanamycin resistant bacteria within a bacterial population can be used to make recommendations for how to remediate a contamination problem. 7. Develop skills with using serial dilution method to determine bacterial concentration. 8. Develop skills in sterile technique and bacterial plating. 9. Develop skills to determine the frequency of antibiotic resistant bacteria in a culture. 10. Develop skills with Polymerase Chain Reaction (PCR) as a method for amplifying a particular DNA sequence. 11. Develop skills with agarose gel electrophoresis for determining molecular weights of DNA fragments generated by PCR. 12. Understand how these processes are used to solve real world epidemiological problems. Performance Objectives: The student will be able to: 1. Determine the concentration of bacteria in a culture using serial dilution methods. 2. Determine the frequency of antibiotic resistant bacteria within a bacterial population based on the number of colonies growing on plates with and without the antibiotic and the dilutions used to plate the bacteria. 3. Use sterile technique for transferring and plating bacteria. 4. Prepare and run an agarose gel for DNA gel electrophoresis. 5. Explain how DNA gel electrophoresis separates DNA fragments by their size. 6. Describe how particular genes carried on plasmids can be used to trace bacterial contamination. 7. Describe the PCR process for amplifying DNA, including the role of the Taq polymerase and primers. 8. Explain how plasmids replicate and are transferred among bacteria in nature. 9. Explain the selective advantage of antibiotic resistance in nature, and when bacteria growing in nature benefit from having plasmids containing antibiotic resistance genes 10. Synthesize knowledge about antibiotic resistance, natural selection, and laboratory techniques used in the experiment to solve a real world epidemiological problem.
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2 Week 1 Overarching Research Problem: Using Genes for Antibiotic Resistance to Trace Source(s) of Bacterial Contamination Create a heading in notebook: “Research Problem: Using Genes for Antibiotic Resistance to
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