D2O Adaptation Day 42

Results:

  • D2O yeast (gen 33) – 3.128 at 24h
  • D2O yeast (gen 34) – 0.726 at 0h

I finally got my DDW order so today I did a starter culture of yeast in DDW YPD inoculating a culture from glycerol stock. I also did my usual 9ml of D2O YPD with 1ml of D2O YPD yeast culture (from the previous generation). Tomorrow I hope I can compare the 24h growth of nonadapted yeast in D2O vs adapted D2O yeast. And then the day after that will be a time trial.

Also I made some D2O YPD plates today. So once the starter culture grows I can also compare the growth of colonies on solid media. That should be a fun experiment. Here is my method:

  1. I already had D2O YPD, and I have agarose instead of agar. I added 0.8g of agarose to 40ml of D2O YPD.
  2. I heated the mixture on a hotplate/stirrer to incorporate the agarose. Normally this step is done in an autoclave, but I can’t use that because I don’t want to add more H2O than necessary to the D2O (d-exchange).
  3. Pour 15-20ml per plate. I made enough for 2 plates.