In preparing the gel, a flask was used to weigh agarose. To this was added 80 ml of 1x TAE. Then, the flask with its contents was placed in a microwave to facilitate thorough mixing for a period of 1 min. The contents were removed from the microwave and carefully mixed again, then placed on the bench and allowed to cool. Upon cooling, 3 Î¼l of ethidium was added. As the gel cooled, masking tape was used to securely tape the two open ends of a gel boat. The desirable comb was placed in the boat, after which it was placed on the bench. 1x TAE was used to fill up the gel tank apparatus, making sure that the electrodes faced to the right. Upon the setting and solidification of the gel, the tape and the comb were both carefully removed and the gel placed in the tank. The tank was filled continuously until the level of the buffer had exceeded that of the gel.
During the process of loading and running the gel, 6x DNA loading dye was used to mix the DNA samples. The mixed DNA samples was then loaded into the wells
The lid was slid into the apparatus, and the black and red cords plugged into the powerpack. The gel was run for between 1 and 3 hours at approximately 80-110V. The Gel Doc System was used to visualize the gel.
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