This protocol will make 1 liters. Adjust to your needs if necessary.
Final concentration of this medium is 1/2 MSMO, 5mM MES (pH5.8), 0.8% (or 0.7%) agar.
Put 700 ml Milli-Q H2O in a beaker
Weight 2.2 g of MSMO (in the 4°C freezer) and add it to the water (agitating)
Add 1.0g of MES (2-(N-morpholino)ethanesulfonic acid)
Adjust pH to 5.8 (adding KOH)
Fill with Milli-Q water to exactly 1 liters
Put 8 g (for vertical plate) or 7 g (for horizontal plate) of Agar in 1 clean glass bottle
Autoclave for 30 min
Let cool, add antibiotic to your convenience and pour into plates
Big round plates take about 50 ml of media
Square plates take 25 ml of media (for horizontal plate) or 40 ml of media (for vertical plate).