Hello again from Baylor University!
It’s hard to believe it’s already week five of the REU program in chemistry! We just completed half of the program, and this week was filled with lots of research successes, setbacks, and other fun things outside the lab.
First off, this week was filled with many more hours in the lab. Last week, I mentioned I had performed my transformation and got bacteria colonies, so I was hoping that I had successfully made some mutations in the DnaB helicase of E.colli. This week was spent screening the colonies that grew for possible successes. We started by doing a genomic prep to purify the genomic DNA from each of the colonies we wanted to screen (I screened about six colonies from each of the six mutations] for a total of 36 colonies screened this week). We then amplified the EcDnaB gene in each of those genomes and finally digested the PCR product with restriction enzymes that should only cut the DNA if we had a successful mutation.
Although we were able to successfully amplify EcDnaB in all of the colonies, we were not able to get any restriction enzyme activity with any of the colonies. We aren’t really sure why this is happening since the literature quotes a 65 percent success rate in this procedure, so I’ll be spending week six working on an explanation. I plan to work mostly with our positive control this week. If I can get the positive control to work properly, then there should be no reason for my other mutations not to work. If that’s the case, I’ll simply repeat the procedure and screen more colonies. It could be the literature value isn’t correct and the mutation rate is much lower than 65 percent. If that’s the case, a larger population should help me get the desired mutations.
Outside the lab this week, we attended a Fourth of July celebration that the city of Waco and Baylor put on. This took place on Baylor’s campus, and thus we were able to walk over and enjoy the food trucks and watch the fireworks. We also spent Sunday cheering on the U.S. women’s national soccer team as they defeated Japan in a 5-2 victory in the FIFA World Cup. Although the focus of the REU is on research and developing future graduate students, it’s nice that we’re also afforded the opportunity to enjoy the Baylor campus and events put on by the city of Waco.
I look forward to week six and furthering my research. I will also spend this week working on a presentation that we all will give on our research at the conclusion of our time here. With the help of my PI, Dr. Trakselis, I am learning how to use new software to create interesting graphics that will serve as great visual aids on posters and presentations to come!