These media solutions will work for propagating M. smegmatis and mycobacteriophages. There are only 3 solutions that you need!
MP Buffer (Mycobacteriophage Buffer)
Solution: Concentration: For 500ml
Tris (pH 8) 50 mM
NaCl 150 mM 4.35g
MgCl2 10 mM .47g
CaCl2 2 mM .11g
Middlebrook 7H9 Liquid Medium (For growing liquid cultures of M. smegmatis)
Distilled water 1000 mL
Middlebrook 7H9 powder (Difco catalog no. 0713-01-7) 4.7 g
Glycerol 5 mL
Autoclave at 15 lb/in2 for 20 min. Ready to use once cooled.
Middlebrook 7H9 Solid Medium (For growing M. smegmatis)
Distilled water 1000 mL
Middlebrook 7H9 powder (Difco catalog no. 0713-01-7) 4.7 g
Bacto-Agar
Glycerol 5 mL
Autoclave at 15 lb/in2 for 20 min. Let cool until the solution can be touched-to-the cheek without squinting (55°C), then pour your plates (25 mL/petri dish is ideal). Let cool on bench, stirring for 30 min.
Pour plates. Some use a 25 mL pipette, I prefer to be more expedient and pour directly from the flask.
Middlebrook Noble Top Agar (When attempting to make plates that contain mycobacteriophage plaques.)
Use 0.6% Noble Agar (Difco catalog no. 0142-17-0) with 0.5% 7h9 powder (Difco Catalog no. 0713-01-7). I.E., use 3.0 g Noble Agar and 2.5 g 7h9 powder per 500 mL water. Autoclave the solution, and leave in a 55°C bath until ready to pour (the bottle can just be touched to your cheek without wincing). Remember, if too hot, the solution will kill your microbes! (If too cold, the solution solidifies and can't be used unless it has been put in the microwave for a minute.)