Finally the starter cultures worked! But not really in the way that I expected. The stuff that was two days old grew and none of the new cultures that I started yesterday grew. Well at least I have something to work with today. Here is my experimental setup:
- Setup of five samples:
- 9ml of DI YPD + 1ml of DI starter culture
- 9ml of DDW YPD + 1ml of DDW starter culture
- 7ml of DDW YPD + 2ml of D2O YPD + 1ml of D2O starter culture (30% D2O sample)
- 4ml of DDW YPD + 5ml of D2O YPD + 1ml of D2O starter culture (60% D2O sample)
- 9ml of D2O YPD + 1ml of D2O starter culture (99% D2O sample)
- Measuring:
- 400ul of each starter culture in semi-micro cuvettes – to measuring starting absorbance
- 400ul of each timed culture (the samples to be measured over time) in semi-micro cuvettes – to measure the time=0 value (should be close to 1/10 of the starting measurements)
- repeat B every hour
- Blanking:
- Before each hourly measurement you need to blank the nanodrop
- Blank the DI samples with DI YPD (no yeast added)
- Blank the DDW samples with DDW YPD
- Blank the 99% D2O, 30% D2O, and 60% D2O samples with D2O YPD