Pglo Transformation Lab Report

The process of transformation is used often to evolve or change a cell by implanting new genes which can safely alter the genes already located in the organism we are trying to change. In this lab, the goal was to transform E coli. to glow in the dark, or in other words to make transform it and give it the ability to be bioluminescent. Bioluminescence is one of nature’s most useful and stylish abilities. In many species the ability to create a glow based off the DNA located in the cell is used in a variety of ways- anything from the glow of a firefly to attract a mate to the way an anglerfish uses this ability to attract prey with the light atop its head. This bioluminescence is only possible through a specific DNA which contains this ability-

On the second day after these colonies were grown, the lab begun by labeling two test tubes -DNA and +DNA, adding the transformation solution to to each, and placing it on ice. Then the procedure called for the use of a sterile loop to pick up a colony of E coli and add it to each test tube, then after E coli was in both test tubes, the pGLO DNA was placed in ONLY the +DNA test tube. After 10 minutes on ice, and after the 4 plates are labeled each to LB -DNA, LB/amp -DNA, LB/amp +DNA, and LB/amp/ara +DNA respectively, the test tubes are heat shocked to loosen the DNA cell membrane to allow pGLO to slip in and code within the DNA. After more time in the ice bath, the tubes are taken out and set at room temperature with the LB broth placed in both test tubes. Then the dishes LB/amp +DNA and LB/amp/ara +DNA were spread with the +DNA solution, and the dishes LB -DNA, and LB/amp -DNA were spread with the -DNA solution. Finally, the 4 plates were incubated once more before the third day, in which we observed our

I believe this for a multitude of reasons, the biggest being the fact that the lack of any growth in either +DNA solution filled petri dishes means that while a transformation might have occurred, the lack of any growth means it was impossible to prove whether or not it did in fact occur. Another piece of evidence is the surplus of failed experiments in our class, despite even people who finished way before the class ended faced no growth in their petri dishes or no glow at all. This suggests the possibility of a less developed experiment equipment, or even the possibility that the parameters we set up were incorrect or maybe that the efficiency rate was lower than it could be. The possibility can be traced back further that the only growth that did occur was in the LB -DNA dish for our group, which suggests the the -DNA substance was not tampered with and grew quite well in a lawn growth pattern. The +DNA substance however grew nowhere, suggesting it had been tampered with. The room for human error is evident here, as we did have a moment where we almost added the solution without the LB broth, and we did add some to the LB/amp +DNA petri dish by accident. Yet, the LB/amp/ara +DNA petri dish was not tampered with before the LB broth was added, meaning the error or lack of proper DNA transformation occurred before this step was used. So, the