Bio110AntibioticResistanceLabManual2013_

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Unformatted text preview: anamycin resistance gene is present at each chicken farm. Create a heading in your notebook: “Experiment Part D: Preparing Gels and Electrophoresis Apparatus” Procedure Preparing Your Gel Solution: Record any intentional or unintentional changes in procedure in your science notebook 1. Make sure your gel apparatus is properly assembled. 2. Weigh 300 mg (or .3 g) of agarose and place in a 125 ml Erlenmeyer flask. 3. Measure 30 mLs of 1X TAE buffer using a graduated cylinder and pour this into your flask. 4. Gently swirl to mix the agarose into the buffer. 5. Place the flask in the microwave and microwave for approximately 35 seconds. 6. Remove the flask using the heat protective glove (bright orange) and gently swirl. Look at your gel solution and make sure that it is clear and there are no particles of agarose remaining. If there are particles, microwave an additional 5 seconds and check again. 7. Let the gel solution cool for approximately 2 minutes. 8. After cooling, put on a pair of disposable gloves. Add 1 µl of ethidium bromide dye (orange solution) to the molten agarose and gently swirl. Dispose the pipet tip in a RED biohazard bag. (Caution! This dye is a mutagen. You must wear disposable gloves when handling the dye and any gel containing this dye.) 9 Casting the Gel: Record any intentional or unintentional changes in procedure in your science notebook 1. It is essential that the dams are wedged snugly into their slots in the gel rig. If the dams are not snug enough, the molten agarose will leak out before it hardens and you will have to clean out the unit and start over. 2. Once you are sure the dams are snug, slowly pour the molten agarose into the gel tray. 3. Remove any big bubbles that might form, especially around the comb, with a yellow tip. 4. Dispose the yellow tip in the RED biohazard bags on the tables. 5. The agarose should harden within 10 minutes; once ready, it will appear slightly opaque. Carefully remove the dams from the unit and return t...
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This note was uploaded on 02/06/2014 for the course BIO 110 taught by Professor Hass during the Fall '11 term at Pennsylvania State University, University Park.

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