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Bacteriological Analytical Manual (BAM) Media M189: Cellobiose-Colistin (CC) Agar

May 2004 

Solution 1

Peptone10 g
Beef extract5 g
NaCl20 g
1000X Dye stock solution*1 ml
Agar15 g
Distilled water900 ml

 

Adjust to pH 7.6. Boil to dissolve agar. Cool to 48-55°C.


 

*1000X Dye stock solution

Bromthymol blue4.0 g
Cresol red4.0 g
Ethanol, 95%100 ml

 

For consistent medium color, use dye solution rather than repeatedly weighing out dry dyes. Dissolve dyes in ethanol for 4% (w/v) stock solution. Using 1 ml of this solution per liter of mCPC agar gives 40 mg bromthymol blue and 40 mg cresol red per liter.


 

Solution 2

Cellobiose10 g
Colistin400,000 units
Distilled water100 ml

 

Dissolve cellobiose in distilled water by heating gently. Cool. Add antibiotics and filter sterilize.

 

Add Solution 2 to cooled Solution 1, mix, and dispense into petri dishes. Final color, dark green to green-brown.

NOTE: This medium, like TCBS, is very inhibitory and does not require autoclaving. Medium may be stored 2 weeks at refrigeration temperatures.

 

Storage:limit: 2 wksTemperature: 4°C