S-STEM Scholar Madison Jackson's Blog

  Intro My time this week was dedicated specifically to exposing deinococcus sonorensis to various levels of ultraviolet radiation and to view the impact on its ability to survive radiation based on what type of media sonorensis is grown on. Specifically, the medias of TGY and R2A were tested and observed for growth. Methods Visual diagram of process shown above 3 biological reps each of TGY and R2A, grow sonorensis in broth of respective mediums measure OD600 value for each sample, normalize to 1 expose 50 uL of sample to radiation (2500, 3000, 3500, 4000, 5000*100uJ/cm^2) for 5 mins return exposed sample to 450uL of pure media, vortex to incorporate inoculate onto plate of respective media wait and observe for growth Results & Discussion Growth was seen on none of the R2A plates, but was present on the 3000 and 5000 TGY plates It should also be noted that while trying to normalize the OD600 values before moving on to exposure, the sonorensis grown in R2A was significantly eas...

  Intro Again, the focus of this week was sonorensis. It was officially decided this week that, considering the upcoming ANAS meeting, we would fully pivot to being copletely dedicated to the sonorensis project as opposed to trying to complete the two separate projects. Though we did start a run of RNA isolation on Tuesday, it was decided to be the last one for now, and was ended early. Part of the week was also spent discussing our poster, who would be responsible for which sections, and the general layout of the poster (what to include, images where, color scheme, etc.).  Methods Take 100 μl of sample and expose to 200mJ/M^2 of UV radiation for treatment period.  After exposure take the 100μl and add it to 900μl of TGY Incubate at 30 °C for 30 minutes before moving on to RNA isolation  RNA isolation Resuspend fresh or frozen pellet in 800 μl RNA lysis buffer and transfer the mixture to a ZR bashingbead lysis tube Secure the tube on a bead beater fitted with a 2ml ...

  Intro As the ANAS meeting approaches, the AMC project is largely taking the backseat, and we pivoted towards the sonorensis characterization tests this week. Much of the time this week was spent making various medias needed for the tests, and then giving them their first run. There was a run of RNA isolation done on e. coli at the very end of the week. Methods Skim milk media was made using the recipe on the bottle (100g/L skim milk powder + 15g/L of agar) diluted down to 100mL SIM media was made using the recipe given on the bottle (30g/L SIM media powder + 15g/L agar) diluted to 100mL Urease plated were inoculated with sonorensis, as well as a positive and negative control (staph. epi and bacillus respectively) E coli UV exposure and RNA isolation as follows: Take 100 μl of sample and expose to 700mJ/M^2 of UV radiation for treatment period (2 min& 4 min)  After exposure take the 100μl and add it to 400μl of TGY Incubate at 30 °C for 30 minutes before moving on to RNA ...

  Introduction     This week, most of our time was dedicated to the sonorensis project, as well as a discussion into the basics of qPCR.   Most of the time spent with sonorensis was discussing what further tests need to be completed and/or done again; the time spent on the AMC project was spent doing another run of RNA isolation. Methods Exposure Take 100 μl of sample and expose to 200mJ/M^2 of UV radiation for treatment period. (5min &10 min in this case)  After exposure take the 100μl and add it to 400μl of TGY Incubate at 30 °C for 30 minutes before moving on to RNA isolation  RNA isolation Resuspend fresh or frozen pellet in 800 μl RNA lysis buffer and transfer the mixture to a ZR bashingbead lysis tube Secure the tube on a bead beater fitted with a 2ml tube hold assembly and process. 1 min on/2 min ice repeated 10 times centrifuge the tube for one minute to pellet debris transfer up to 40 0 μl of the cleared supernatant into a Zymo-Spin IICG colum...