I would like to find a method of increasing the biomass of my D2O cultures because my current method is not yielding enough protein. I would like to also minimize the amount of H2O in my culture.
My current methodology is a small scale growth of 3mL with 75% D2O and LB grown for approximately 9hrs. This culture is then used to seed a 20mL 100% D2O and M9 culture overnight and this entire culture is used to seed a 1L D2O and M9 culture.
Thanks for your time.