In addition to the Day 33 daily adaptation I’m going to run an hourly experiment to see if the yeast has adapted or evolved or changed. I’ll be looking for a growth rate that is faster than in previous trials, and more comparable to yeast growth in DDW YPD. Here is my setup:
- Prepare the YPD in test tubes, I’m doing final volumes of 10ml:
- 95% DDW – 9ml of DDW YPD
- 35% D2O – 6ml of DDW YPD, 3ml of D2O YPD
- 50% D2O – 4.5ml of DDW YPD, 4.5ml of D2O YPD
- 65% D2O – 3ml of DDW YPD, 6ml of D2O YPD
- 99% D2O – 9ml of D2O YPD
- Add yeast:
- add 1ml of yeast grown in 50% D2O YPD to all test tubes except the 99% D2O YPD
- add 1ml of yeast grown in 99% D2O YPD to the 99% D2O YPD test tube
- Incubate at 30C
- Measure in Nanodrop hourly starting at t=0. The starting absorbance values are:
- 95% DDW – 0.511
- 35% D2O – 0.512
- 50% D2O – 0.505
- 65% D2O – 0.634
- 99% D2O – 0.439