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U.S. Department of Health and Human Services

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(BAM) Media M35: Cefsulodin-Irgasan Novobiocin (CIN) Agar or Yersinia Selective Agar (YSA)

January 2001

  1. 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

     
  2. Irgasan (Ciba-Geigy) solution
    Irgasan 0.40% in 95% ethanol 1 ml

    May be stored at -20°C up to 4 weeks.
     

  3. Desoxycholate solution
    Sodium desoxycholate 0.5 g
    Distilled water 200 ml

    Bring to boil with stirring; cool to 50-55°C.


  4. Sodium hydroxide, [5 N] 1 ml
  5. Neutral red, [3 mg/ml] 10 ml
  6. Crystal violet, [0.1 mg/ml] 10 ml
  7. Cefsulodin (Abbott Labs), [1.5 mg/ml] 10 ml
  8. Novobiocin, [0.25 mg/ml] 10 ml
  9. 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.