Nearing the End

During the previous week, I picked transformed E. coli colonies that were mentioned in the previous blog post. I put these colonies into media and incubated them at a temperature that would allow them to grow at optimal reproductive capacity. After approximately 24 hours of growth, I created a pellet of the cells in a centrifuge and performed a plasmid preparation procedure on the pellet. This procedure resulted in purified plasmids, on which I then performed a restriction digestion. After this I performed gel electrophoresis on the restriction digestion in order to see if the reaction worked and found out that it did. In the following day, I extracted this restriction product from the gel, and executed a procedure on it in order to purify the DNA in the product. In the future (most likely the beginning of fall semester) I will do procedures to incorporate this purified DNA product into a viral genome that will be used to infect E. coli cells in a process called transduction. During transduction, a viral vector will incorporate it’s genome, that includes the hyper mutable region of interest, into E. coli cells. Once this is done, I will begin performing the same mutation rate procedures done on H. pylori on these E. coli cells.