Yeast Growth in D2O, DDW, and DI YPD: Setup

Yesterday I created a starter colony of yeast from the lyophilized yeast that I ordered from Today I’m doing growth in different water types and checking the growth every hour just like the e. coli experiments. Here is my setup:

  1. Make YPD broth with DDW and D2O.
    1. the bottles of both come in 100g quantities which means there is 90-ish ml of D2O and 100-ish ml of DDW
    2. I had 91ml of D2O –> 4.55g of YPD powder
    3. 100ml of DDW –> 5g of YPD powder
  2. suspend powder in water types and filter via syringe. DO NOT AUTOCLAVE
    1. the reason is because of deuterium exchange. The powder is ultrapure and the water is pretty pure itself, so filtering should be just fine.
  3. To make diluted cultures:
    1. I have 4 samples: 1) DI water 2) DDW 3) D2O and 4) 50% D2O in 50% DI water
    2. Put 9ml of each water type into a test tube (except for the 50/50 mix which should be 5ml of d2o and 4ml of di water)
    3. Add 1ml of starter culture (which is made from DI water)
    4. put in incubator
  4. take time points every hour in semi-micro cuvettes using nanodrop. I fill them with 0.5ml of each culture.