September 11, 2020
By: Abby Maruska and Zoe Rivard
This semester we have been getting back on track since the Covid-19 pandemic has hit. Our current goals because of COVID-19 regulations and limited lab time, are to finish sequencing our gene, SCN3B, and successfully complete RNA isolation. We have had lots of ups and downs in the research completed so far, some being; our primer dilutions were messed up, our gel hasn’t shown the results we wanted, and our RNA isolation did not have a strong purity. Since we have gotten back into lab after many months of being confined to our homes, we want to improve these tests results and keep moving toward our goals.
Since coming back from last spring, the first protocol we performed was a PCR and then a gel electrophoresis. We first made new primer dilutions because we had run out of the ones from last semester. Abby and I followed our notes and made the new 0.5M primers. We chose to run our gel with only CDNA and H2O, no beta actin controls. After imaging our gel, we realized we had a problem. On our gel there was no bands in the CDNA, which there are supposed to be amplified at our gene’s certain band. We concluded that this problem must be from our primer that were just made, as we haven’t had this problem last semester. Now our next step is to remake our 0.5µM primer along with a 1µM primer and run another PCR and gel.
While working on sequencing our gene, we also have been running an RNA isolation protocol. This protocol is longer than others and has lots of small, important steps that can’t be missed. Eventually we will be doing this same protocol with a brain sample, but since we are just getting used to this process, we ran it with a liver sample. The most difficult part of RNA isolation is homogenizing the liver sample quickly and completely without the RNA denaturing. A problem we ran into early on was that our liver sample was not homogenized fully, this led to our final RNA not having enough concentration. Because of this we are going to perform another RNA isolation with liver samples before moving to the brain samples.
COVID-19 had put a pause on our research, but not anymore. Being able to get back on campus and start up our research again has been awesome, even though our test results have not gone as planned. Reworking and conducting experiments to get better results means we get more practice and time in lab. Overall, we’ve had many challenges so far, but Zoe and I are putting in the work and aiming for our goals.
