S-STEM Scholar Alex Hugenberg-Cox Blog

      This week was prepping for the first LexA E.coli  qPCR, meaning we first had to make sure that the UV limits were correct then perform an RNA isolation. The beginning of the week was spend busy with biotech bootcamp for the first few days, but after that i got started with the UV exposure to ensure that the levels that we expected to see significant impact wouldn't kill them. For this i did a simple experiment by exposing them at three different levels of UV radiance (50, 60 and 70 mJ/cm^2) for two different times (1 and 2 minutes. All of the samples were exposed then plated alongside a control, and grown till the next day. All of the plates had growth so we were able to use the highest levels and times at 70  mJ/cm^2 for two minutes during our RNA isolation.      The next day I did the RNA isolation, diluting the samples grown that morning (due to the doubling time of E.coli) to an OD of 1 +/- .05, then exposing 100 μl and after letting them recover in 900μl of LB broth gett

     This semester I am still continuing work with the AMC (activated methyl cycle). This project has been continued from the 2023 spring semester, through the summer and fall semesters. While the experiment has changed throughout the semesters the core of the project has remained the same: looking at the gene expression change after oxidative stress in a species within the Deinococcus genus.       This semester I am taking a step back from the main AMC project, and instead focusing on a different gene (metK) within the cycle. While this is still very similar to the main project is covers a different gene and does not go The goal of this project is to develop a verified procedure using E.coli , see metK's response to UV exposure within Deinococcus caeni , and have a poster for this seasons conferences.  The reason we are looking into E.coli at the beginning instead of going straight into  D.caeni is so that we can verify/validate our qPCR techniques on a on-experimental organism a