Jekielek_Week 6 Reflection
I really appreciated the very engaged discussion we had this week. It was really helpful to have a walk through with this paper as I quickly got lost with the different strains, the primer optimization, and then the different methods used. There were a lot of parts to the paper that became clearer to me throughout the discussion and the presentation by Shane, Dara, and Emma. I had not anticipated doing any PCR as part of my project, but it was really good to learn the application of it for essentially verifying the materials and methods being used. It really clarified for me the differences between PCR and qPCR and how they can be used in tandem for crossvalidation. I had no idea what a primer dimer was, so that was interesting to learn how impactful they can be to your results. I didn’t think to ask this question during class, but can you identify primer dimers if you were only using qPCR? My intuition says no. I’m still not fully clear on the multiplexing. I’m also not clear why we can’t use a method like reverse transcriptase PCR to identify different ontologies in a species. My understanding is that to determine life stages we need to utilize RNA…so why can we not use this method for teasing that out??
Great paper, great discussion. I’m now generating my own data and figures, so this paper came at a perfect time.