E. Coli Cells Lab Report
Erica Osorio
5057497
Christian Roque and Rogerlio
The Mechanisms by which E.Coli Cells Developed Immunities toward Ampicillin due to Plasmid and DNA Consumption
U34
Abstract During the ampicillin experiment the ability to transform cells to make them adaptable to their environment was studied. The E.coli bacterial cell was used in order to observe how its DNA was able to change and develop immunity towards ampicillin. In order for this change to occur the use of several plasmids was needed. The plasmids used in this experiment were pUC18 and the lux plasmid. The E.coli cells that developed the resistance to ampicillin by the use of the pUC18 plasmid will be able to grow and divide in an ampicillin environment. Those that transformed successfully with the lux plasmid will be able to multiply in an environment that is subjected to lux and will also glow. In order to perform the experiment the cells had to be made competent before the start of the experiment in order to enhance the uptake of plasmid DNA. The competent cells were then incubated with plasmid DNA. Afterward, the cells that had taken up plasmid DNA and …show more content…
Use a sterile micropipette to add 5 µL of control plasmid into both the “C” and “lux” tubes. Going back to the tube with competent cells, add 70 µL of the cells into each of the two test tubes. Mix these solutions and store both tubes in an ice bath for 15 minutes. During this time obtain an additional tube and add 35 µL of competent cells into this tube. This will become the “NP” tube or no plasmid. Take all the test tubes and transfer them into a water bath preheated to 37°C and let them sit for about 5 minutes. Add 275 µL nutrient broth to the control and lux tubes and add 150 µL of nutrient broth into the no plasmid tube. Let the tubes sit in this bath for about 45
5057497
Christian Roque and Rogerlio
The Mechanisms by which E.Coli Cells Developed Immunities toward Ampicillin due to Plasmid and DNA Consumption
U34
Abstract During the ampicillin experiment the ability to transform cells to make them adaptable to their environment was studied. The E.coli bacterial cell was used in order to observe how its DNA was able to change and develop immunity towards ampicillin. In order for this change to occur the use of several plasmids was needed. The plasmids used in this experiment were pUC18 and the lux plasmid. The E.coli cells that developed the resistance to ampicillin by the use of the pUC18 plasmid will be able to grow and divide in an ampicillin environment. Those that transformed successfully with the lux plasmid will be able to multiply in an environment that is subjected to lux and will also glow. In order to perform the experiment the cells had to be made competent before the start of the experiment in order to enhance the uptake of plasmid DNA. The competent cells were then incubated with plasmid DNA. Afterward, the cells that had taken up plasmid DNA and …show more content…
Use a sterile micropipette to add 5 µL of control plasmid into both the “C” and “lux” tubes. Going back to the tube with competent cells, add 70 µL of the cells into each of the two test tubes. Mix these solutions and store both tubes in an ice bath for 15 minutes. During this time obtain an additional tube and add 35 µL of competent cells into this tube. This will become the “NP” tube or no plasmid. Take all the test tubes and transfer them into a water bath preheated to 37°C and let them sit for about 5 minutes. Add 275 µL nutrient broth to the control and lux tubes and add 150 µL of nutrient broth into the no plasmid tube. Let the tubes sit in this bath for about 45