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key 12 - KEY Bio 205L EXERCISE 12 Prep Sheet for LABORATORY...

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KEY: Bio 205L EXERCISE 12 Prep Sheet for LABORATORY PRACTICAL EXAM Procedures and Techniques learned during this laboratory: If the same technique is used in multiple sections of a lab, it does not need to be listed multiple times. ( For the lab practical exam, be sure you know how to properly perform each technique). Section 11b.2.: Isolation of Plasmid DNA from a Bacterium. a. Alkaline Lysis DNA Mini-Prep procedure 1. Centrifuge overnight bacterial cultures at 3000 rpm. b. What is the function of each of the following steps? 1. Addition of STE buffer to the cell pellets. 2. Addition of GTE buffer to the cell pellets. 3. Addition of Alkaline SDS solution to the plasmid prep. 4. Addition of Potassium Acetate solution to the plasmid prep. 5. Centrifugation following the Potassium acetate step. 6. What is present in the supernatant following the centrifugation and what is in the pellet? c. Know how to precipitate DNA using ethanol, and explain why this is done.
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