Characterization Tests

 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

  1. Take 100μl of sample and expose to 200mJ/M^2 of UV radiation for treatment period. 

  2. After exposure take the 100μl and add it to 900μl of TGY

  3. Incubate at 30°C for 30 minutes before moving on to RNA isolation 

RNA isolation

  1. Resuspend fresh or frozen pellet in 800μl RNA lysis buffer and transfer the mixture to a ZR bashingbead lysis tube

  2. 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

  3. centrifuge the tube for one minute to pellet debris

RNA isolation was stopped here, and then frozen.

  • SIM media inoculated
  • inoculated both types of casein plates
  • inoculated 1 flask of TGY & 1 flask of R2A with deinococcus sonorensis for UV exposure later
  • poured citrate plates for later inoculation

Results & Discussion
Many of the tests that had to be redone were incoulated at the very end of the week, so results will not be seen until next week. The sporulation tests were decided to be inconclusive, as our results did not line up completely with either a negative or positive result and will be done again. Other than that, a good chunk of time this week was spent talking about our upcoming poster and the division of labor. The very basic plan is shown here:
In general, every person in our group has at least one section that they will be responsible for with our group leader Alex functioning as overseer of everything, as well as having her own sections she's completely responsible for.

Conclusion
Moving forward we are continuing with the characterization tests, as well as getting photos for our poster. With abstracts due soon, it is possible we may be helping with that though only in small ways like phrasing and such.

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