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Pglo Transformation Lab Report

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Our experiment followed the genetic transformation of the bacteria E. Coli. Genetic transformation is the dynamic take-up of free DNA by bacterial cells and the heritable fuse of its hereditary data (Lorenz and Wackernagel 1994). In this case, the E. Coli was combined with green fluorescent protein to allow it to glow in the dark. Other components, such as arabinose sugar, uniquely composed pGLO, and the antibiotic ampicillin resistance, are all combined to create the fluorescence with a UV light source. Green fluorescent protein is what will cause the fluorescence in the bacteria and must be blended with arabinose, a sugar that will stimulate the glow. It is through the process of heat shock, we were able to genetically transform the bacteria …show more content…

In the presence of ampicillin, E. Coli growth will not occur. Therefore, in this experiment, bacterial growth on the plate indicates the uptake of the ampicillin resistant gene into the genetic code of E. Coli. If pGLO, LB broth, ampicillin resistance, and arabinose sugar are added, then the bacteria will glow. In one of the four plates that include all of those component, fluorescence will be observed. The purpose of this experiment is to demonstrate the genetic transformation that occurs in the E. Coli bacteria. The importance of this experiment stems from E. Coli’s factors. E. Coli is known to be a predominant coliform and they cannot endure living in non-enteric conditions (Megraw and Farkas 1993). Since this bacteria can form diseases in the body, it is critical to distinguish …show more content…

We used a micropipetter to place calcium chloride into both microcentrifuge tubes. The use of the transformation solution, CaCl2, is to amplify the plasma membranes permeability. Both of these tubes will have entered ice for a short period of time. After, with a sterile loop tool, a single group of bacteria from a beginner plate will have been placed in these tubes. Only in the +pGLO tube, pGLO Plasmid DNA will be submerged into with a sterile loop and then placed into the tube. The tubes will have been placed in the ice for an extended time and eventually it is time to place them into warmer water to create the heat shock treatment. This treatment is there to expand the plasma membranes permeability and it boosts the rate of a complete transformation. While these actions occurred, 4 plates will be needed and labeled to separate which compounds go into which plate. The plates will feature one with +pGLO, LB, and ampicillin, the second has +pGLO, LB, ampicillin, and arabinose, the third contains -pGLO, LB, and ampicillin, and the final had -pGLO and LB. The use of the micropipette helped place each of these arrangements into the plates and is then spread equally around with a sterile loop. Tubes and sterile loops have been disposed into a red autoclave bag. The plates were placed into a 37 degree celsius incubator and our results will be known next class (Weedman

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First, label one micro centrifuge tube +pGLO and another –pGLO. Using a sterile transfer pipet, transfer 250µl of competent cells (E. coli + CaCl2) into each tube and place them in crushed ice. Examine the pGLO plasmid DNA solution with the UV light and note your observations. Pipet 10µl of pGLO plasmid into the +pGLO tube and mix, close and return it to the ice rack. Do NOT add plasmid DNA to the –pGLO tube.

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The purpose of this lab was to perform a procedure known as genetic transformation which allowed us to genetically engineer E. Coli to be ampicillin resistance. Before the lab we expected that lysogeny broth and minus DNA will have growth but no glow. The lysogeny broth, ampicillin, and

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