1. Make up the medium according to the directions and prepare plates containing
approximately 20ml of a basal layer of TSC or SFP Agar containing egg yolk.
2. Prepare 0.1ml aliquots of a suitable series of dilutions of the homogenised test sample and
spread over the surface of the basal layer using a sterile swab.Overlay with an additional 10ml of
egg yolk free TSC or SFP Agar. Cultures which are not overlaid with agar are unlikely to
grow as black colonies.
3.Incubate the plates at 35°C for 18-24 hours with an anaerobic Gas Generating Kit (BR0038) in a gas-jar.
Alternatively, use AnaeroGen (AN0025 or AN0035). AnaeroGen does not require the addition of
water or a catalyst.
4. Alternatively, pour-plates using approximately 25ml per plate of TSC or SFP Agar containing
egg yolk may be prepared using 1ml aliquots of a suitable series of dilutions of the homogenised
test sample. Mix the plates well before the agar gels. With this technique, lecithinase activity of
Clostridium perfringens colonies is more difficult to see. Clostridium perfringens colonies may be seen
as large, black (2-4mm diameter) colonies within the depth of the agar.