February 20th, 2014
Lab report 4
This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.
This lab was meant to reveal the variations of different
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We then used a transfer pipet to add 250μL of cold calcium chloride to each tube. During all the steps that these tubes weren’t needed they were placed on ice. We then used a sterile plastic inoculating loop to grab E. coli colonies from the starter plate to the “+ plasmid” tube. This loop was then inserted inside the calcium chloride solution and spin to ensure that the colonies were transferred. We then suspended the cells to ensure an homogeneous mixture. Furthermore we transferred a mass of cells to the – plasmid tube and suspend like we did with the “+ plasmid”. Our team used another inoculating loop to add one loop full of plasmid DNA to the + plasmid tube. The “+ plasmid” tube was then added to ice for 15 minutes. During that time we labeled our plates. After the incubation we put the tubes in 42°C water for 90 seconds of heat shock. 250 μL of Luria broth was added to each tube. The tubes needed a 5-15 minutes recovery time at room temperature. The cells were then removed from each tube and spread on the plates. To spread the cells we used plastic beads, which uniformly spread the cells by shaking them across the plate. The process was repeated for each plate. Finally the plates were incubated at 37°C for 24-36 hours and then we could find results.
Results My groups results:
(-)Wild Cells (JM101 strain)