• I want to build and maintain a library of frozen bacterial cultures.
• Is it possible to make the concentration of glycerol in my growth medium 10%, before the addition of cultures (Grown on streak plates).
• The first organism I'm trying is ATCC 9895. I am unsure how the growth of the bacteria will be affected by the properties of the glycerol at this concentration.

I have a feeling that the osmotic stress caused by the 10% glycerol could possibly delay or even prevent the bacteria from reaching log phase. I will find out pretty quickly if it will grow, but my main concern is viability of the frozen cultures in the long term. If anyone has experience with this scenario, please let me know!