Science is difficult. I said that I expected this coming into the program, and indeed I have been proven to have anticipated correctly. Oddly enough, however, I came into this program expecting the theory behind the science to be the most difficult part, not only in terms of understanding (in my case, for example) mechanistically how genes can interact with one another in a way that produces specific regulatory pattern and the implications of those regulatory patterns, but also in terms of understanding how the data that we hope to obtain will shed light on these questions.
As it turns out, this expectation was wrong. Understanding the theory behind my research has proven far more manageable than what has proven to be a far more tricky, and sometimes cruel, adversary: project design.
At first, my project seemed easy enough to plan. I knew where I was starting, and I could see and understand plainly enough the general steps that I needed to take in order to reach the goal.
Step 1: Take this promoter and put it into this plasmid.
Step 2: Take this gene and insert it next to the promoter in the same plasmid.
Step 3: Repeat steps 1 and 2 with different combinations of promoters and plasmids. What I did
Seemed like a straightforward enough task to complete to me. But little did I know. It turns out that completing step 1 alone is much more easily said than done. Not only are there a barrage of procedures required to make the plasmid (each of which introduces innumerable opportunities for error) but there are also a barrage of procedures required to make sure that the end product is the one I intended. After all, I have to make sure that I end with plasmid A instead of G, X, V, L, or any other possible mistake.
Having to plan and carry these steps out carefully enough to be confident that I am actually producing the plasmids I need has challenged me more than any other aspect of my project. I can’t count the number of times at each step of the process that my mentor has asked me “But what about this?” or “Did you account for this?” Often, the answer is “uhhhh…no” and it’s back to the drawing board. As I’ve undergone this cycle many, many times over these six weeks, it’s often felt like I’m taking one step forward, glancing over my shoulder at least three times to make sure that I haven’t messed up, and, realizing I have, taking half a step back. That’s a definite woe.
But don’t misinterpret. Making it through that process has been one of the most rewarding parts of this program. Being able to say “Ok, this is where I am now and I need to get to this point” and then being able to make an outline of procedures, one that efficiently accomplishes the task while making the proper and needed security checks, has been an incredibly rewarding and empowering. I’ve yet to master the art (in fact I just found out that my plasmids may not function properly given that my original template plasmid seems to have a faulty promoter), but I’ve enjoyed learning. Despite the frustration of having to redo several steps along the way, I still find myself excited by the idea of doing it right.
