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
- Dissolve ingredients in small beaker and adjust pH to 5.6
- Filter sterilise solution using 0.22 μm filter.
- Store in refrigerator.
Solution B (Aqueous Bromothymol Blue)
- Bromothymol Blue 0.4 g
- 0.01N NaOH 64 mL
- Distilled water 36 mL
- Dissolve the Bromothymol Blue in the NaOH
- 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
- Autoclave to 121C for 15 minutes, cool to 48C.
- For plates add 100 mL of the filtered solution A and mix. Dispense as plates.