Zabulionis Unit Notes

Zabulionis Unit Notes - Bio 2290G Winter 2010 Rotation 3...

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Bio 2290G: Winter 2010: Zabulionis Unit: Rm 330 NCB Rotation 3: Session 1: Welcome: Transformation in E. coli Welcome to the first lab session of the Zabulionis Unit of Biology 2290G - Scientific Methods in Biology. Beginnings are very important; during this session we'll get to know one another and this unit better; we’ll have a look at what is in store for the next three weeks. It is much better to be proactive than reactive — read, understand, and be prepared for each session. In the laboratory sessions following the first two, you will design and carry out experiments to answer a question of your choice regarding the biology of bacteria and plasmids. Some of the biological context for your experiments will be presented today. This laboratory session also introduces some conceptual background and technical skills through exercises involving transformation of E. coli bacteria. Objectives: Review set up of Lab book according to guidelines in the Resource Manual (pg 90 - 91) and discuss Unit style. Prepare your protocol for the steps involved in the transformation of E. coli . Micro-pipet usage (please don’t wreck them!!!) Safety: fire extinguisher, shower, gas shut- off, eye wash station, EMERGENCY. Transform E. coli cells with plasmid DNA.
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Basic Transformation Protocol: E.coli JM101 cells on pick plate add to cold calcium chloride ice divide into 2 tubes add TE add plasmid ice 45 0 C heat pulse ice add LB broth 37 0 C incubation plate incubate When recording a protocol in your lab book—can someone pick up your lab book and do the experiment exactly the way you did it without your guidance???
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Safety: Lab coats, long pants, and glasses required for bench work. NO open-toed shoes. Long hair tied back. Beware of hazards associated with Bunsen burners, handling of bacteria etc. . BE AWARE OF WHAT YOU ARE DOING!!! Background Prep: Antibiotics, Aseptic technique (Pouring plates, Transfer techniques, Pipetting tips), Plasmids, Safety in the Laboratory, Scientific Nomenclature, and Transformation background material in Resource Manual. Read the “Practical” file on WebCT so you are aware of the skills we will be testing you in Session 6 and will be learning in this lab period. Assignment(s): “Exit Stamp” for Transformation Protocol in Lab Book.
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Session 2: Transformation Background The first experiment will provide context for a discussion of basic principles underlying transformation and antibiotic resistance in bacteria. The relevance and importance of controls will also be discussed along with proper data recording procedures. Most of the next session is devoted to designing experiments.
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Zabulionis Unit Notes - Bio 2290G Winter 2010 Rotation 3...

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