I started a little over a week ago at my lab. I haven’t received my own project yet as I am shadowing a post-doc in the lab, learning the techniques that are employed when conducting actual research projects.
To start, I was asked to transvect three different plasmids into E. coli and then extract these plasmids from the bacteria, linearize the samples, and obtain the linearized DNA to save for future use.
My first attempt in transvecting these bacterial cells with the plasmids didn’t go as smoothly as planned; after regrowing the bacterial culture with the transvected cells a second time, the plasmids were successfully extracted from the cells and concentrated so that all I had in test tubes was plasmid DNA. With this circular DNA, I got to linearize them using restriction enzymes and then proceeded to perform PCR to allow the DNA to amplify. After PCR, the products were run on a gel to check and make sure that the samples indeed were linearized properly and that the bands were of the right length.
Then, when all this was done, I expected to start on a new project, to learn a new skill. But I was wrong. Yes, I learned a new skill but I was not given anything new to work on. Instead, the post-doc showed me how to, after imaging the gel, extract the DNA from the gel and proceed to obtain this linearized DNA only. Even though I used a pre-made kit in order to extract my DNA from the gel, it was fascinating to see how all this is done. I never thought that I could do anything with the gel and the DNA bands that are produced. In the past, after imaging, the gel was discarded and that was it. The DNA was never saved. But I have always wondered, how do labs get such large quantities of linearized DNA of a specific vector if the only way that a vector can be amplified is by transvecting bacteria? Well, I have my answer.
I love lab. I love learning, how hands-on the entire process is, and how everything that I have learned in my past classes are actually coming alive and are applicable. I am glad I chose the major I did. I absolutely love it. And I can’t wait to learn more as the year goes on.
[Sorry for the lack of coherency in some parts. It is a pretty stressful week with several quizzes, a midterm, and a midterm paper due for me. Hopefully, I will begin to write more after all this is over and write with more finesse and eloquence. Thanks for reading!]