Boston!

I am privileged to have had the opportunity to travel to Boston for 2  weeks during Summer 2024 to attend my in-person lab requirements.  Prior to this trip, I had never been on the East Coast, and this was such a wonderful experience in exploring my home campus and bonding with  my peers, whom I had only worked with virtually thus far. 

In class, we were introduced to many important isolation methods and  techniques for analysis, including gel electrophoresis, PCR, Elisa,  Western Blot, HIC Column Chromatography, HPLC, BCA assay,  restriction digest, and DNA/protein extractions. We had the opportunity  to establish our own prokaryotic and eukaryotic cell cultures. We  performed transformation on our E. coli cells, and transfection with our  HEK297T cells to introduce foreign DNA tagged with a GFP marker for  identification. We performed serial dilutions and stainings with  Coomassie and Ponceau, learned how to interpret a chromatogram, we  annotated vectors, and identified appropriate primers. 

We also learned how to read a UV-Visible light absorption spectrum  obtained by means of measuring optical density to determine DNA and  protein concentrations. 

 This was the pride of our research- we spent two weeks culturing  HEK293t cells, which are an immortalized human embryonic kidney cell  line. I had not idea how tedious it was to culture eukaryotic cells. We had  to check on them twice a day and change media frequently. There were a few times we weren't sure if our line was going to make it! We measured  pH, CO2 uptake, Temperature, nutrient concentrations, lactic acid... and this was a highly simplified experiment- I can only imagine the challenges  scientists encounter in the field! We generated pGlo in E. coli cultures, then  isolated the plasmids containing the fluorescent marker with HIC Columns.  Two days before these images were taken, we introduced the pGlo plasmids  to our  eukaryotic cells by means of transient transfection. In transient  transfection, there is no incorporation of the foreign genetic material from  the plasmids into the host cell genome, so over time as the cells continue to  divide and the transfected cells die off, the glow dims. We were the only  group in the lab whose cells were successfully transfected!!!