How to create long term stock for Bacillus subtilis? I am trying to create a long-term stock for Bacillus subtilis (Bacterial glycerol stocks). I have looked online for guides and step-by-step tutorials and I found that they don't say where the sample originally comes from. In my case, I am using **Bacillus subtilis subsp. subtilis (ATCC® 23857™)** pure from the supplier bottle. What is the proper procedure to create a long-term stock for Bacillus subtilis using this product?

I worked with _Bacillus subtilis_ WB800 and just used 50% glycerol / 50% LB. I imagine the same would work fine with wild-type. I would prepare the glycerol/LB mixture and autoclave it then create the stocks from log-phase (approximately) plate cultures. This method was sufficient for my application (protein expression).