D2O Adaptation Day 44 Time Trial Setup

Two days ago I inoculated a starter culture of yeast in DDW YPD, and let that grow for 48h. Yesterday, I inoculated another culture from the previously mentioned culture to ensure that my yeast for today’s experiment would be in the log-phase of growth. Then I took a sample of my normal D2O yeast and compared the growth of this yeast against all the others and made a new sample of the yeast grown in DDW YPD to be grown in D2O. This would be to finally determine if the growth of regular yeast in D2O is obviously different than the yeast that I’ve been incubating in D2O for the past 44 days. Here is my setup:

  1. Put 9ml of YPD in a test tube. I want 4 samples, with two samples being DDW YPD and the other two being D2O YPD.
  2. Add 1ml of culture to each sample. One sample has 24h incubated DDW YPD yeast, one has 48h incubated DDW YPD yeast (see above), a third has generation 36 D2O YPD yeast, and the final sample is 48h DDW YPD yeast in D2O.
  3. Incubate at 30C.
  4. Record absorbance via nanodrop hourly.

The main motivation (as I said above) is to compare the growth of my 44 day incubated D2O yeast vs regular yeast grown in D2O. Normally the yeast in D2O starts at a really low absorbance value for example ~0.020 compared to yeast in DDW (ie ~1.000) or D2O adapted (~0.700). I wanted to start with a much higher cell count to see if the growth would be any different.

The other motivation is to compare the growth of yeast after 24h of incubation vs 48h of incubation. I was worried that they time trials I’ve been doing haven’t been using yeast in log-phase growth, and if their growths are comparable I know that I’m not.