Yesterday I set up starter cultures for this experiment using D2O, DDW, and DI water as the basis for the liquid media. Today I’m running the time trial experiment again, but this time with a bit more of a spectrum in water amounts. Here is the setup:
- I am using 6 samples: DI water, DDW, 30% D2O mixed with DDW, 60% D2O mixed with DDW, 90% D2O mixed with DDW, and 99%D2O.
- For the mixes:
- add D2O, add DDW, add 1ml starter culture from DDW (1:10 dilution).
- Example: 30% D2O in DDW is 3ml D2O, 6ml DDW, and 1ml DDW starter culture
- For the other samples:
- DI: 9ml of DI YPD, 1ml of DI starter culture
- DDW: 9ml of DDW YPD, 1ml of DDW starter culture
- 99% D2O: 9ml of D2O YPD, 1ml of D2O starter culture
It should be noted that the 99% D2O starter culture was almost completely translucent. By this I mean it looked like there was no growth in the sample at all. Results that I’ll post later (when the experiment is complete) will show that this is indeed true, with an absorbance of 0.521 (compared to ~2.1 in both DI and DDW starter cultures, 4x the cell growth!).
I’ll be taking growth readings every hour via the nanodrop in micro cuvettes (til about hour 4) and then switching over to semi-micro cuvettes (because I’ll run out of micro cuvettes).
Love,
Ant