S-STEM Scholar Alex Hugenberg-Cox Blog

      This week we went back to our own oxidative stress experiment. After a long discussion about our lack of results when using Hydrogen peroxide as a stressor, we decided to switch our stressor from that to Sodium Hypochlorite (NaOCl). Even though we never got to test our results from the exposure where we introduced catalase, Chad was certain they would yield results no different than any of the tests we had done before. Instead it was decided to go with Sodium  Hypochlorite, from bleach.      Before we could start our actual experiment, we would need to see what percentage of sodium hypochlorite would be ideal. We would be using agar plates, and mixing the cells with different dilutions of NaOCl and inoculating. The plates would than incubate for 48 hours before establishing how each dilution percentage effected the cells. We started by doing a dilution from household bleach at roughly 8% sodium hypochlorite, diluting down to 5.25% originally before doing further dilutions. (From

      This week was very uneventful. Chad had put our project on hold for the week, and we instead spent our time helping Lani both with final poster prep and the initial stages of the DNA extraction project. While they had started the project over the summer, the samples they had were in the wrong elution and would need to be redone.      We spent the week doing graham stains, inoculating broths and plates, making gels and refilling more pipette tip boxes than one would thing possible.      All in all we did a lot of busy work that to us wasn't much but helped others when they were to busy to do it themselves.      The conference itself also went well. This was the Arizona Nevada Academy of Science conference. There was a wide variety of posters that were all very interesting and covered a wide array of topics. Two of the ones I spent a good deal of time at was a poster that studied bite forces of sharks compared to body mass, and one that studied plants to hear ultrasonic noises

       This week we ran our RNA samples from last week, so i got to make and run a RNA gel for the first time.      Monday Deja transformed a small amount of our T1 RNA sample into cDNA using a new kit. Our usual kit was gone, so in the mean time we tried a small amount of our sample with a new kit that had different steps to see if it would work. The only way to see if it had worked was to run qPCR, so that's what we did Tuesday. Unfortunately it did not work, so we would need to wait for our usual kit to get it, but we still had the rest of the samples.      Wednesday I taught Lizzie and Iditi, to people in the lab who had never done the RNA isolation procedure, how to do it. It was good for me to sit and run through all the steps, and explain each one and make sure it was being done correctly and precisely. It helped me gain a better understanding of the procedure, and what different steps did.      Thursday was mainly doing a few things to help Lani out with her upcoming poster

      After reviewing the original paper our project was based on at the end of last week, we decided to try and edit our hydrogen peroxide exposure protocols in order to get a better result. This mostly meant adding catalase at the end of our incubation period, along with some other minor changes.      We started the week of with some inoculations and graham stains, and spent a good amount of time preparing for a biotech program booth that we would be setting up later that day.      Wednesday we finally got to the new hydrogen peroxide exposure. We started by diluting down 11.5M H2O2 down to 200mM. We also diluted down our six groups of cells all down to an OD close to 1.  After pelleting all of our cells, we removed the supernatant, and resuspended all of the cells in 200μl of PBS.  We than re-pelleted, and added 200μl of H2O2 to our test cells, and 200μl of PBS to our control cells. Next we incubated the cells for 30 minutes at 30 °C. After we immediately added 100 μl of catalase to