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L-Canavanine Glycine Bromothymol Blue (CGB) Agar

For differentiation of C. neoformans and C. gattii (Kwon-Chung et al. 1982).

Solution A

  • Glycine Univar 10 g
  • KH2PO4 1 g
  • MgSO4 1 g
  • Thiamine HCl 1 mg
  • L-canavanine sulphate 30 mg
  • Distilled water 100 mL
  1. Dissolve ingredients in small beaker and adjust pH to 5.6
  2. Filter sterilise solution using 0.22 μm filter.
  3. Store in refrigerator.

Solution B (Aqueous Bromothymol Blue)

  • Bromothymol Blue 0.4 g
  • 0.01N NaOH 64 mL
  • Distilled water 36 mL
  1. Dissolve the Bromothymol Blue in the NaOH
  2. Add to the water.

To prepare medium (1 litre for plates)

  • Distilled water 880 mL
  • Solution B 20 mL
  • Bacto agar (BD 214010) 20 g
  1. Autoclave to 121C for 15 minutes, cool to 48C.
  2. For plates add 100 mL of the filtered solution A and mix. Dispense as plates.
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