Prepare nutrient broth as recommended. Dispense 99 ml portions to 170 ml bottles. Autoclave 15 min at 121°C. Cool to room temperature before use.
Dissolve 0.1 g lysozyme in 65 ml sterile 0.0l N HCl. Heat to boiling for 20 min. Dilute to 100 ml with sterile 0.0l N HCl. Alternatively, dissolve 0.1 g lysozyme in 100 ml distilled water. Sterilize by filtration through 0.45 µm membrane. Test for sterility before use. Add 1 ml lysozyme solution to 99 ml nutrient broth. Mix and dispense 2.5 ml portions to sterile 13 × 100 mm tubes.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.