Going into the program, I certainly expected experiments to completely fail. I expected them to return completely unexpected results. I expected them to be unidentifiably wrong in some way. But what I did not expect is for the experiments to tease me with how close they were to what I wanted.
For much of the summer, my major goal has been to confirm the identity of several plasmids. In order to do so, the plasmids need to first be isolated from bacteria and resuspended in solution. Then restriction enzymes are used to cut up the DNA into single strands that can be visualized on gels in order to see if the expected bands appear.
The first step of isolating the DNA started off well enough. I was able to produce a solution of at least 0.1 micrograms of DNA per microliter. The problem was that two of the plasmids just wouldn’t surpass this threshold. One only reached 0.09 micrograms per microliter, while the other only reached 0.07 micrograms per microliter. I attempted each at least three times, and each time, it was so close, but so far. It was not a complete failure, but just not enough success to warrant using the solution in future experiments.
Also, when using the restriction enzymes to confirm the identity of the plasmids, it seemed as if only a fraction of each run would work, even though there doesn’t seem to be a reason one would work, while another wouldn’t. I started with ten plasmids. Two were confirmed during the first run. Four on the next run. Two during the third attempt. Only one was confirmed during the fourth attempt. The final confirmation required streaking, culturing, purifying, and digesting all over again in. It reminded me of Zeno’s dichotomy paradox that only allowed a weary runner to travel half the whole distance, then half of the fraction remaining, and so on, never reaching the finishing line.
Despite taking much longer than anticipated to confirm the plasmids, there was a great amount of satisfaction after accomplishing something that took that much patience. Additionally, it was much faster to see the application of the labor as two of the plasmids were transfected into HEK 293 cells and shown to express in vibrant reds and greens.
What I have definitely learned through this experience is that the best accomplishments cannot occur suddenly or without trial. Being able to do something immediately, without effort or thought just breeds complacency with the result. The best joys of research have been at the end of periods of exacerbation.
While I am quite confident that my project will produce results within the next week and a half, I am hesitant to admit so since it might jinx the effort. Just as I have learned to trust but verify results, I have also learned to hope for a good outcome, but be prepared for an unexpected result.