S-STEM Scholar Melanie Castaneda's Blog

  This week we were able to begin working with D. roseus to try to find its minimum inhibitory concentration. In order to move through our list of species quicker, we had to begin working on several different bacteria at a time rather than waiting to wrap up a species before moving on to the next. D. roseus was the bacteria with an appropriate amount of growth that would allow us to work with it.  Performing MIC Protocol on D. roseus  Pipette 5 ml of TGY into each test tube 13 x 100 mm test tubes with metal caps  7 for each of the three antibiotics that will be used  21 test tubes total Autoclave to sterilize tubes and TGY Take the OD600 value of bacteria  Want it between 0.80 - 1.00 If the value is higher, dilute it with TGY until it is within our desired range  Our sample of D. roseus was at 1.76 so we diluted and the new value was 0.9 Once TGY has been autoclaved and cooled, remove appropriate amount of TGY per tube  Amounts are shown in the ta...

  Monday, February 27, 2023  MIC Testing with D. aquaticus  To continue our MIC testing on D. aquaticus, we prepared 8 test tubes with metal caps with 2.5 mL of TGY with 0.5% agar. They were autoclaved for 30 minutes in order to be ready to be used for soft agar plates. These soft agar plates will be used to test the higher concentrations of kanamycin on D. aquaticus. The concentrations will be 1000 ng/mL, 2000 ng/mL, and 3000 ng/mL, and our negative control which will not include kanamycin. Each concentration will have a duplicate which brings our total plate count to eight.  Inoculating D. indicus We prepared to begin working with D. indicus. Since we plan on working with all of the Deinococcus species that we have available in the lab, we decided to continue our MIC testing with D. indicus. We took the OD600 value of our culture that we had growing in broth from 2/9/2023. A range that we aim for when it comes to OD600 values is anywhere between 0.80 - 1.00. The v...