Pipetting RNA for qPCR analysis

Author: Drew Vander Leest        Major: Biology

My research was a part of a NASA initiative called the Planetary Protection Act. Essentially it is to ensure that other planets aren’t contaminated from biological matter from Earth during interplanetary missions. I specifically studied the expression of two proteins of a psychrophilic bacteria that could hypothetically survive Mars’ conditions based on its resilience to the cold and salty brines that it thrives in. The proteins of interest were a cold-shock protein and a phosphate transporter with several membrane-spanning domains that were tested under different solutions of chaotropic salts and freezing temperatures. The end results and interpretation of the expression levels can pave the way for understanding how chaotropic salts can interfere with lipid bilayers and biological processes like transcription and translation.

I was first introduced to this topic by Dr. Mack Ivey, who was my research mentor. I had no idea what chaotropicity (the focus of my project) even meant prior to researching its effect. So, in a grand sense, I was able to learn a lot about this topic as it was central to the project. To clarify, it is how some solutes can affect the entropy and different phases of water (like exerting the hydrophobic effect and rearranging solvation layers). Beyond understanding the scope of my project, I learned a great deal about general lab procedures that enable more precise results. This includes proper aseptic techniques, good pipetting form, and how to perform routine tests like PCR and gel electrophoresis. I found that I really enjoyed studying how to do these techniques and procedures. It was interesting to actually do the same work that was used to obtain the knowledge I was learning in a lot of my classes. So being able to apply the scientific method in my work gave me an admirable perspective of everything I was currently learning simultaneously.

I am grateful to be a part of Dr. Ivey’s lab because it gave me a lot of agency in the research process. I was looking at several labs in the biology department, and I was familiar with Dr. Ivey because of his involvement in the premedical honors society and as a premedical advisor. I viewed his work on Clostridium and asked him if he had any openings to work on that specific project. He gave me the opportunity to work on that original project, but also mentioned another NASA project with an anaerobic bacteria that is tested under Mars’ conditions. I thought that it was a very intriguing topic so I went with the latter one instead. Dr. Ivey was a great mentor as he helped design the experiment with the topic of interest. He helped me develop my thesis and grant proposal, and he managed the lab well. I also worked with Sergio Mora, a PhD candidate who was working on a parallel project. He helped me with technical lab abilities and taught me a lot about how to work in a lab.

I did encounter some issues during my research tenure. The ongoing pandemic affected my research project in several different ways. The most impacting consequence was the fact that I was unable to work in the lab from the middle of March to the middle of October. This severely affected how the research was able to be conducted. Because of the limited amount of time available to complete the project, it had to be modified. There was only enough time to do a single qPCR analysis of the protein expression from the samples. I also focused on qualitative research after having to make some adjustments.

Because of my recent graduation, I plan to work as a research assistant at a clinical research institution before I begin my medical school career.