355. CLOSTRIDIUM PYG MEDIUM

Trypticase peptone 5.0 g

Peptone from meat (pepsin-digested) 5.0 g

Yeast extract 10.0 g

Salt solution (see below) 40.0 ml

Na-resazurin solution (0.1% w/v) 0.5 ml

L-Cysteine-HCl × H₂O 0.5 g

Na₂CO₃ 2.5 g

D-Glucose 5.0 g

Distilled water 1000.0 ml

Salt solution:

CaCl₂ × 2 H₂O 0.25 g

MgSO₄ × 7 H₂O 0.5 g

K₂HPO₄ 1.0 g

KH₂PO₄ 1.0 g

NaHCO₃ 10.0 g

NaCl 2.0 g

Distilled water 1000.0 ml

Dissolve ingredients (except cysteine, carbonate and glucose) and sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Add cysteine, then dispense under 80% N₂ and 20% CO₂ gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose from a sterile anoxic stock solution prepared under 100% N₂ gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Adjust pH of complete medium to 7.0, if necessary.

Updated 11.07.2021