It’s almost the end….
We are coming up to the end of the semester here in the Williams lab.
Larissa, Michelle, Nancy, and I are all working diligently to finish our work this semester. These last weeks before Thanksgiving break will be our last opportunity to collect data for thesis. The entire thesis experience has been very challenging but very rewarding. Over the semester, I have been improving our protocol for Chromatin Immunoprecipitation with zebrafish embryos. We previously were analyzing the result with PCR and gel electrophoresis that yielded semi-quantitative results, but now we are utilizing quantitative PCR to produce more quantitative results. Following the first couple of trials with qPCR as the follow up analysis we noticed that there was still a lot of nonspecific binding occurring during the immunoprecipitation step, and the wash buffers were not reducing that binding enough to feel confident about the results. This led to a number of changes to the protocol including modifying the incubation time with the antibody, the wash buffers following the IP, and adding a pre-blocking step before the IP. I am working on the pre-blocking solution now. With each change, we have seen promising results for a more specified ChIP protocol in zebrafish.
For last week and this week, Nancy and Michelle have been presenting their data that they have gathered over the semester. It’s been interesting to see their specific projects develop as well. Michelle and Nancy have been able to gather some amazing images and data on the effects of oxidative stress chemicals on vertebrate morphology and gene expression. Michelle’s images with the confocal microscope are beautiful and its incredible the clarity to which we can now visualize microscopic structures within animals. Nancy has also gathered a wealth of data on the Nrf1 response to oxidative stress. While I am excited to present my own data, I also wish I had more time to continue the project and being determining the effects of dioxin on expression of different Nrf genes. I have learned so much this semester and Larissa has been a wonderful mentor throughout the process. I am excited to continue this work in the future on my path into the research field of molecular biology.