Now that the lab is clean, I’m going to collect my thoughts and write a plan to follow. While the first two experiments were unsuccessful, there were some interesting results that can’t be disregarded. But in the pursuit of science more results are required! So let’s get this party started…
D2O adaptation: Based on the paper I read yesterday, it seems that a slow adaptation is the right way to go. Yeast grown in pure D2O seemed to endure too much stress to adapt and the adaptation might be way slower compared to a progressive adaptation plan. So each week I’ll increase the amount of D2O by 20% starting with 20% D2O YPD:
- 20% D2O
- 40% D2O
- 60% D2O
- 80% D2O
- 100% D2O
- Every Monday I plan on running a time trial experiment comparing the growth of yeast in DI YPD to the increased D2O YPD concentration and to the previous D2O concentration.
- Every Friday I’ll run a time trial of the yeast growth in that week’s D2O concentration.
- Every day, yeast will be provided new medium for growth and daily nanodrop readings will be recorded to analyze the 24 hour growth. Also a microscope analysis will also be done to ensure there is no contamination.
- Weekly glycerol stocks will be created in case of contamination.
- Agar plates of each D2O concentration will be created to analyze colony morphology. Inoculation onto solid media will take place each week, and right now I’m not sure when the best day to do this is, but I’m thinking Wednesdays might be best.