Science Fail Monday: The Benchtop ExtractionPosted: 05 04, 2015
Protein purification can be a lengthy process that requires patience, perseverance, and, at times, creativity. The purification of a protein that is susceptible to degradation and exhibits poor solubility provides extra challenges.
I had the pleasure of working with one such protein. The purification procedure calls for three different affinity columns and numerous incubation steps. In between each affinity column, fractions containing the protein are identified via gel electrophoresis and either concentrated to a smaller volume, or incubated with proteases to remove affinity tags. During hour thirteen on day five, I was preparing to filter my sample before loading it onto the last column. Somehow the sample ended up on the lab bench instead of the filter, thus inspiring an unconventional technique known as the “benchtop extraction”. I pipetted the protein off the benchtop and loaded it onto the final column. Surprisingly, this technique has yet to be published in Nature… perhaps due to the devastatingly low yield.
In hindsight, the story is amusing; however, at the time, I felt completely defeated. How often do days, and even weeks, of hard work yield no results? But I suppose that’s the nature of research: if everything worked perfectly the first time, a Ph.D. would not be held in such high regard, not to mention, most diseases would already have a cure.
As scientists, we aspire to make a significant contribution to our respective fields. To do so, we must investigate the unknown. While the “bench-top extraction” could have easily been avoided, all scientists are faced with setbacks and failures. Some weeks, it seems like Murphy’s Law governs our lives. But those weeks make the days when we finally get some promising data all the more rewarding. Being a good scientist first and foremost requires determination and persistence. And when you inevitably hit the metaphoric wall, remember that you are not alone, and that everything works out in the end.
by Rachel Cohen
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This article by Rachel Cohen was co-published on the SWAC Blog, The Pipettepen: http://swac.web.unc.edu/thepipettepen/