Transformation Lab Report
Abstract (Jenna):
In the transformation of E. coli bacteria the purpose was to transform DNA segments into the E. coli to make it glow just like a firefly. Transformation efficiency was to be calculate to determine if using E. coli in vaccines would create a stronger more potent vaccine against an illness or if another bacteria would be more efficient when compared.
Introduction (Kyle):
Discussion (Jenna): Add in background information about the lab
To transform E. coli bacteria it was necessary for all group members to have goggles and gloves on at all time to reduce the risk of contamination from students and ingesting the bacteria. Agar solution was essential for the growing of bacteria colonies. It provided a base for the bacteria to flourish into colonies that were needed for later parts in the lab. Small amount of calcium chloride was also used to mix with the E. coli in different test tubes and a majority of the time for the last half of the experiment the E. coli was in test tubes. Because of the small amounts that were being used there was no need to use larger equipment such as a beaker for mixing substances together.
The lab was carried out over a period of four days for two reasons: to allow enough incubation time for the E. coli to colonize and there are a couple of places in the experiment that there is a waiting period. The pre-lab was carried out over two of those days. With a break in between days two and three gave enough time for the bacteria to colonize. The design of the overall experiment was very well planned out. It allowed a time for the bacteria, but also enabled the students to not have to rush through the lab and end with strange results. Planning out the days and what needed to be accomplished always lead into the next lab day that we had. Because there were a couple of moments in the lab that had waiting periods built-in, the lab was extended longer than a usual lab. That gave time to prepare for other portions of the lab.
At the end of the experiment the group’s working assumption was correct that by adding in the segment of DNA to allow the E. coli to glow worked. The glow wasn’t quite as strong as what was hoped for, but there were other groups that had E. coli that glowed very well and the group was able to observe.
Methods (Lauren):
To start the experiment, two tubes were filled with ice-cold calcium chloride and then put on ice. Using a plastic inoculating loop, transfer E. coli colonies into the tube marked with a +. Mix these using a pipet until the calcium chloride is a milky white. Return the tube to ice and repeat with the tube marked with a -. Add plasma DNA to the tube marked with a +. Leave both tubes in ice for 15 minutes. After the 15 minutes are up, place the tubes in a hot water bath for 90 seconds. After this time, place the tubes back into the ice for one minute. Add Luria broth to both tubes and mix by tapping the sides of the tube. After the LB is mixed in, pour the liquid inside of the test tube on the plates, matching each sign to each other. Use beads to spread out the mixture on the plate. Wrap the plates up with tape and incubate them for a little over a day.
Results/Data (Kyle):
Conclusion (Lauren):
In this experiment, we learned about bacterial transformation. One important part of this lab were the plasmids. Plasmids are exploited by bacteria in order for the bacteria to grow. This information is vital to our current project. We now understand what controls need to be used in order to transform E. coli in our project. We also now understand vaccines and diseases, so we can incorporate that into the final product. This lab showed us how to properly describe the experiment in a video, which is going to be the final product for the project.