Day 4 - The Start of Experimentation

For the last day of the first week of my J-Term, I had finally started to work in the lab itself.

I arrived at Memorial Hermann in the morning, and made my way to the same lab once again. I was then greeted by the doctor that I would be shadowing and working with today. We both walked over to the cell culture room, so that she could explain to me the ongoing experiment that she was working on. She was working on cell cultures of course, and specifically, how CD73 plays a role in turning Adenosine into ATP, and how silencing that same CD73 changes the process as a whole.
After a brief explanation of what we would be doing for that morning, for the remainder of this morning I watched her work on the controls that she needed for this experiment, as she already finished with the actual cell cultures a few days before, on Monday. She also explained what she was using for the controls, like the media, lipofectamine, etc. After she finished, we let them sit for 5 hours, and I went off to lunch

After lunch, she started doing PCR, or Polymerase chain reaction. She was doing PCR to detect the RNA and DNA of the mice, and certain tissues of the mice. I watched her use the snippets of the mice tails she had cut, to put into mastermix to genotype the mice. Then, she had put the tubes into a machine called a "thermal cycler" which would then denature the DNA by heat, splitting the DNA from a double-strand to two single strands. Then it would anneal the primers that were added to the DNA mixture, to the target. This means that the primer would bond. The last step would be extension, which is when more base pairs are added. For the sets of PCR that she had done, each one needed an average 33-35 cycles in the thermal cycler.

After going through both of those experiments, I can say that this day has really helped me further my understanding of this aspect of medicine, as well as immunology and immunotherapy specifically. I also hope to have more knowledgable experiences in the lab in the next week. After we were finished for the day, I headed home for the weekend.

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