Dissolve 25.50 grams in 1000 ml distilled water. Boil to dissolve the medium completely and sterilize by autoclaving at 15 lbs pressure (121°C) for 15 min, cool it to 42-45 °C and distribute aseptically in petri plates. Ensure complete solidification and aseptically add Antibiotic assay medium No. 1 as seed agar, inoculated with the microbial culture.