Week of 11/18-11/22

This week, we checked the growth of transformation plates from the prior week, but no growth was observed. The control plates showed only pigmented growth, and the plates exposed to UV radiation on normal TGY also did not show any growth. Additionally, rehydrated normal D. rad. cells for the NASA project failed to grow. To address these issues, we planned and researched a new transformation protocol and successfully identified one specifically designed for D. rad. The protocol is as follows: 1. Take 5 mL of fresh D. rad. growth (broth) and transfer it into a 15 mL tube. 2. Add 1 mL of CaCl_2 to achieve a final concentration of 30 mM. 3. Incubate for 80 minutes at 30°C, then transfer 1 mL to a 1.5 mL tube. 4. After incubation, add 10 µL of plasmid and hold on ice for 30 minutes. 5. Dilute each tube with an additional 9 mL of TGY broth and incubate for 18 hours at 30°C. Using this new protocol, we performed three transformations. We also prepared one plate of normal TGY with two samples from two different plasmid extractions and exposed it to 100 µJ for 2 minutes and 30 seconds. The results of this trial of transformations are expected the following Monday.

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