It’s been a productive few weeks at the Williams lab.
With many students staying on campus over some of break, there’s been a lot of progress made in analyzing the embryos we’ve been dosing. Using qPCR, we’ve been looking at the relative expression of a variety of antioxidant genes, and are in the process of comparing the expression across our three time points. We’ve been getting through the usual hoops with qPCR, mostly getting our standard deviations across wells below 0.5. This is to ensure that the change in expression value (Ct) that we see is due to actual changes in expression and not just technical errors such as pipetting. In sum, most of us have come a long way and our results get better with each plate of wells.
Another issue is the constant freezing and thawing of samples, especially our primers and our cDNA. We have so many wells to pipette that we have to be quite careful in how we plan our plates, in order to minimize sample loss through freezing and thawing. In total, we have about 500 wells to pipette correctly, a task that seemed impossible to me in the beginning. Due to some thorough planning and practice, however, I feel confident we’ll all have our plates done soon. The next steps include a refresher and some practice on how to pull needles, as well as injecting embryos with our Nfe2 morpholino oligonucleotide. Other than this extra complication in our procedure, the rest of the techniques ahead remain the same as before, allowing for even more practice in perfecting our lab skills.