Blog Post 3: Inflammatory Lipid: Differences in Male and Female Production

Our research focuses on inflammatory lipid production in mice with a mutated COX2 enzyme, but we are currently in the preliminary stages. In our research, thus far, we have focused on genotyping each litter of mice in order to determine in which cases we were successful in the mutation of the gene. Last semester we finished writing a procedure for the process and at this point, have successfully performed two rounds of genotyping. We have experienced some difficulties in this process. The gel for the first litter we attempted to genotype did not turn out. We then performed gel electrophoresis and PCR again, in attempts to determine where we went wrong in the process. Based on the fact that the second gel produced results after a repeat of PCR, we believe that the error occurred at that step in the process. From that point on we changed the procedure in regards to PCR. We reworked the procedure to include mixing at each step in the PCR process and no longer leave the PCR products over night in the machine to prevent issues within the gel electrophoresis. We also recently experienced issues with our shaking water bath, which is used for DNA extraction from the tail tips. During our most recent round of genotyping, the machine experienced difficulty maintaining a constant shaking rhythm, which is necessary for the DNA extraction. We believe this could have happened due to age or a lack of use. Despite these issues, we were able to successfully genotype our current litter.

This process has been extremely helpful to me in gaining knowledge of what research actually looks like. I’ve almost enjoyed some of the setbacks we’ve had if only because it gave me experience problem solving when our experiment didn’t produce viable results. It can be extremely difficult pinpointing the spots in your research were you’ve gone wrong and I’ve never actually had to go through this process in-depth to fix issues that arise in lab. This process has also helped me gain an understanding of an in-depth research process. Although we have a big picture research question, we have only been able to take the first steps toward this process. While it may seem a small part, the genotyping is a crucial step in the whole process. We cannot start to look at prostaglandin production until we know which mice are actually usable in our study. Lastly, I have enjoyed so much, working with and learning from Dr. Upmacis. She has been such a great mentor, including me in the process of problem solving when issues arise.



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