It’s crazy to think where we started just a few weeks ago. Over the span of this semester, we learned how to transform S. cerevisiae DNA to remove the TEP1 gene, measure growth rates using liquid YPD and a spectrophotometer, optimize paper-based, 3D/PDMS, and deodorant-based analytical devices, and compare growth of different yeast strains. We met obstacles and learned how to overcome them. We learned.
Now, here is our final post, a bitter-sweet moment.
Conclusions
- The results across our different platforms were inconclusive. While some trials showed significant growth differences between Wild-type and TEP1d yeast strains under several conditions, most of our data showed their was an insignificant difference.
- What does this mean? We could attribute the insignificant difference to human error, methodology/design error, or too few trials. OR the results could very well suggest TEP1 does not play a prominent role in S. cerevisiae. TEP1 inhibits the activation of SCH9, a gene responsible for response to osmotic pressure and entrance into the G1 phase from G0. However, SCH9 is also activated by a G protein from a secondary pathway. Perhaps the secondary pathway activates SCH9 more than TEP1 inhibits it. Perhaps the G protein reacts to an uninhibited SCH9 protein pathway and responds by activated the protein less often. Only further investigation could tell.
- Novel analytical platforms (paper and PDMS) have real-world use!
- How so? Paper is cheap. All you need is a wax printer, which can double-function as a daily printer, and you can create hundreds or thousands of analytical devices! This is especially useful in underdeveloped countries or underdeveloped facilities for quickly performing microbial tests. The downfall is paper platforms cannot be reused. However, PDMS can be! While the ingredients for PDMS are expensive, once PDMS molds are created, they can be reused by simply washing them. Furthermore, our design of PDMS use does not require expensive machinery for data analysis, like a spectrophotometer.
- Amateur mistakes at work: failed to take daily OD readings of yeast, use buffers for microtiter plate assays, and solidify pH 2 YPD agar (oops).
- How did this affect our results? Without OD readings, we could not ensure the concentration of our yeast inoculations were consistent. Without buffers, the results of each OD reading of the microtiter plate pH solutions were unreliable. The lack of buffers did not maintain the pH of the yeast growth media and made the results incomparable to the other platforms. Without pH 2 YPD agar solidification, we could not obtain results for comparative study between platforms. Future studies would attempt to correct these mistakes
The Lab Journal 