The first part of my research experience went surprisingly smooth, with few mishaps or questionable results, with the exception of an enzyme or two not working as well as we’d hope. It was fantastic to see my first plasmid I worked with (expressing GP120 and Red Fluorescent Protein) successfully be integrated into the poxvirus genome; some cells in our tissue culture glowed red, confirming the gene’s uptake!
The second plasmid I’ve been working with has required much more trouble-shooting, however. After several days of digesting and ligating pieces of DNA, we finally grew up a culture of E. Coli that we’d transformed with our plasmid (to make much more of it to work with). At first glance on a gel its size looked right. But after doing digests to confirm the genes we desired were present, we quickly realized something had gone wrong. After a few attempts to isolate the plasmid of interest from the overwhelming amount of ‘wrong’ plasmid, we finally decided to take another route. We’d order a synthesized segment of the plasmid to work with, making it more straightforward of a process.
While this setback wasn’t major or devastating, it showed me how valuable troubleshooting and the ability to adapt is to research. Things can quickly take an unexpected turn, and it is good to be prepared to deal with those issues and find a way around them. All in all, I’ve extremely enjoyed my research experience thus far and am looking forward to continuing work on this project!
