(BAM) Media M35: Cefsulodin-Irgasan Novobiocin (CIN) Agar or Yersinia Selective Agar (YSA)
- Basal medium
Special peptone 20 g Yeast extract 2 g Mannitol 20 g Pyruvic acid (Na salt) 2 g NaCl 1 g MgSO4.7H2O (10 mg/ml) 1 ml Agar 12 g Distilled water 756 ml
- Irgasan (Ciba-Geigy) solution
Irgasan 0.40% in 95% ethanol 1 ml
May be stored at -20°C up to 4 weeks.
- Desoxycholate solution
Sodium desoxycholate 0.5 g Distilled water 200 ml
Bring to boil with stirring; cool to 50-55°C.
- Sodium hydroxide, [5 N] 1 ml
- Neutral red, [3 mg/ml] 10 ml
- Crystal violet, [0.1 mg/ml] 10 ml
- Cefsulodin (Abbott Labs), [1.5 mg/ml] 10 ml
- Novobiocin, [0.25 mg/ml] 10 ml
- Strontium chloride, [10%; filter-sterilized] 10 ml
May be stored at -70°C. Thaw to room temperature just before use.
Preparation: Add ingredients for solution A (basal medium) to water and bring to boil with stirring. Cool to about 80°C (10 min in 50°C water bath). Add solution B (Irgasan) and mix well. Cool to 50-55°C. Add solution C (desoxycholate); solution should remain clear. Add solutions D through H. Slowly add solution I with stirring. Adjust pH to 7.4 with 5 N NaOH. Dispense 15-20 ml into each petri dish. Commercially prepared dehydrated Yersinia selective agar (Difco) with supplements may be substituted. Follow manufacturer's instructions for preparation.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.