D2O Adapted E. coli Experimental Replication

Yesterday while writing I realized that the images of the D2O adapted E. coli that I’ve taken were grown on D2O YPD. In an effort to figure out if the morphologies are due to the YPD or the D2O, I’ve decided to redo the experiments on LB agar.

Today I made some D2O and DI LB broth:

  • 1.84g LB in 92ml of D2O
  • 1g LB in 50ml of DI water
  • Filtered broth for sterilization

Then from there I made some LB agar

  • 40ml of liquid D2O LB with 0.8g of agar (2% agar)

I already have solid LB plates with normal water (commercial).

I then incubated E. coli at 37C in liquid media so that I can streak the plates and analyze them. I used 2 different E. coli:

  1. Normal competent cells
  2. Cells from Day 33.
  3. I also had made a separate culture from an unlabeled glycerol stock that I’m pretty sure I made when I finished the experiment. I’ll check it out tomorrow after the sample has developed.