Yeast in DI, DDW, 30%, 60%, and 99% D2O Trial 3: Setup

Yesterday I made starter cultures in D2O, DI water, and DDW. Today I’m doing the time trial experiment and here is the setup:

  1. Setup of five samples:
    1. 9ml of DI YPD + 1ml of DI starter culture
    2. 9ml of DDW YPD + 1ml of DDW starter culture
    3. 6ml of DDW YPD + 3ml of D2O YPD + 1ml of DDW starter culture (30% D2O sample)
    4. 3ml of DDW YPD + 6ml of D2O YPD + 1ml of DDW starter culture (60% D2O sample)
    5. 9ml of D2O YPD + 1ml of D2O starter culture
  2. Measuring:
    1. 400ul of each starter culture in semi-micro cuvettes – to measuring starting absorbance
    2. 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)
    3. repeat B every hour
  3. Blanking:
    1. Before each hourly measurement you need to blank the nanodrop
    2. Blank the DI samples with DI YPD (no yeast added)
    3. Blank the DDW, 30% D2O, and 60% D2O samples with DDW YPD
    4. Blank the D2O samples with D2O YPD