Prepare GC agar base according to the manufacturer s instructions. Autoclave it at 121°C and cool the medium to 80°C. Aseptically, add 5- 7% sheep blood. Mix thoroughly and cool to 45°C to 50°C. Aseptically, add 10.0 ml of sterile supplement solution (see below) to 1L medium. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Supplement solution:

Composition per liter:

Glucose 100.0 g

L-Cysteine.HCl 25.9 g

L-Glutamine 10.0 g

L-Cystine 1.1 g

Adenine 1.0 g

Nicotinamide adenine dinucleotide 0.25 g

Vitamine B12 0.1 g

Thiamine pyrophosphate 0.1 g

Guanine. HCl 0.03 g

Fe(NO3)3.6H2O 0.02 g

p-Aminobenzoic acid 0.013 g

Thiamine.HCl 3.0mg

The supplement solution is commercially available as "IsoVitaleX enrichment"