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