McKinney:7H10

Summary

7H10 agar is selective medium used for the culture of mycobacteria, in particular M. tuberculosis. M. smegmatis is usually grown on LB agar.

Ingredients

• 19g Middlebrook 7H10 powder
• deionized water
• 100mL OADC (Dubos/Middlebrook Oleic Acid Albumin Dextrose Complex)
• 10mL sterile 50% glycerol

Protocol

1. Mix 19g 7H10 powder in 900mL deionized water in a 2L with a magnetic stirbar until fully dissolved. Top up to 1L with deionized water in a graduated cylinder, then return to the flask and stir until well mixed. Cover with aluminum foil and autoclave.
2. Place flasks in a 55°C water bath to cool or cool while stirring with magnetic bar. Prewarm OADC in 37°C water bath.
3. In a containment hood, add 100mL OADC and 10mL 50% glycerol to the flask. Replace foil lids, remove from hood, and stir until well mixed. Pipette 24mL per plate into petri dishes. Keep flasks in 55°C water bath until ready to pour.
4. Leave plates stacked i nthe hood overnight, then store at 4°C.