Materials and Methods Two test tubes labeled pGLO and pGLO were placed into a

Materials and methods two test tubes labeled pglo and

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Materials and Methods: Two test tubes labeled +pGLO and –pGLO were placed into a foam tube rack. Then, a clean pipette was used to transfer 250 μL of (CaCl 2 ), also known as transformation solution, into each of the tubes. The tubes were then sealed and placed on ice for three minutes. After being placed on ice, a colony of E. Coli from a starter plate was picked up using a sterile loop and submerged into the +pGLO solution tube. The loop was twirled around until the entire colony dissolved into the transformation solution. The +pGLO tube was then closed and placed back into the rack in the ice. The procedure was repeated with another new loop for the –pGLO solution. Both tubes were left on ice for three minutes. After those three minutes, a new sterile loop was used in the to pick up a full loop of solution in the pGLO DNA stock tube. It was then mixed into the +pGLO tube and placed back onto the ice with the cap of the tube sealed shut. Both tubes were placed into the rack and then placed on ice for ten minutes. While the tubes were still on ice, five plates were gathered: One Luria broth (LB) plate, two LB/Ampicillin plates, one LB/Arabinose, and one LB/Ampicillin/Arabinose plate. Each plate was then labeled for identification later on. After the ten minutes of the tubes being on ice was completed, they were transferred to a water bath, which was set at 42°C. The tubes were in the warm water bath for exactly 50 seconds. The tubes were then placed back on ice for two minutes. Following the
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incubation, the tubes were placed on the bench top and 250 μL of Luria broth was mixed into each solution. The tubes were then incubated in room temperature for 20 minutes. Using a clean loop for every plate, the film of solution was spread over the surface of a plate containing the LB nutrient agar. These plates were subsequently stacked onto a rack upside down and placed into an incubator set at a temperature of about 37°C. The variables in this procedure would be: the
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