NGM Agar

Reagent	Final concentration	Amount
NaCl	3 mg/mL	3.0 g
Bacto Peptone	2.5 mg/mL	2.5 g
Agar A	17 mg/mL	17 g
MgSO4∗(1 M)	1 μM	1 mL
CaCl2∗(1 M)	1 μM	1 mL
Cholesterol (1 mg/mL)∗∗	0.05 μg/mL	5 mL
Potassium phosphate buffer (PPB)∗∗∗	25 μM	25 mL
ddH2O	N/A	968 mL
Total	N/A	1000 mL

Autoclave media containing ingredients without an asterisk. After making, store plates at 4°C and use within 2 months.

Recipe makes enough for 60 plates (15 mL per plate).

∗Filter-sterilize. Add 1 mL per L agar after autoclaving and cooling.

∗∗Dissolve cholesterol to 1 mg/mL in 100% ethanol. Filter-sterilize. Add 5 mL per L agar after autoclaving and cooling.

∗∗∗ Mix 132 mL of 1 M K₂HPO₄ with 868 mL of 1 M KH₂PO₄ to make PPB. Filter-sterilize and add after autoclaving and cooling.