Stepping away from the high and dry


One of the things I’ve always found striking about biology is the juxtaposition between the clean concepts that we learn in lectures and the microscopic messiness of the physical reality.  The gap between theory and application is large because the multitude of nitpicky details are lost in a high-level explanation. Furthermore, the key concept of the procedure is often not actually visible when performing it. In affinity chromatography it’s easy to miss the actual step where FKBP12 binds to the His-tag, because the whole procedure is just combining different clear liquids with each other and centrifuging them for slightly varying amounts of time. 

A really visceral understanding of this gap is one of the most valuable things that I have gotten out of 20.109, especially since a lot of 7.05’s material lined up really well with the theory behind our Mod 1 experiments. Some salient features of the day-to-day in the wetlab:
  1. Working with countless little tubes of clear liquid and trying to picture what’s going on in there at the molecular level
  2. Not being able to really believe that your tubes contain what you think they do until the data confirms it
  3. Or, as is more often the case, none of your results are what were expected, not even the control, and now you have no idea what went wrong at what point
  4. Really wanting your data to be meaningful since you spent so much time gathering it, but alas a p score is a p score
  5. Losing track of the bigger picture when buried in the minutiae of the protocol, and having to remind yourself to take a step back and remember the reason for each reagent. 
  6. Later, trying to write your report and realizing that you’ve low key forgotten the reasoning and theory behind the actual research question. 

In a weird way I almost appreciate that our experiments didn’t work very well and that our data was so incredibly insignificant. What’s more realistic than failure? Our particular ligands may not have bound in any significant fashion to FKBP12, but in a real lab that would just be the first step of the process. We might have tried a different method for purifying FKBP12, run DSF again, run PPIase again, run a different assay entirely, gone back to to the drawing board completely and picked a new list of ligands. Behind every peer-reviewed paper is countless hours in the lab and more negative results then positive, but that’s all part of the process. 20.109 is all about the journey, and that’s what makes it such a valuable learning experience.  

I'm looking forward to Mod 2, setbacks and (hopefully) successes included!

- Giselle Peng

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