Medium data

Search for medium no. = [272].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

272   

KH2PO4  0.4     g
NH4Cl  0.9     g
MgCl2·6H2O  0.36    g
NaCl  10.0     g
Sodium acetate  0.5     g
Yeast extract (BD-Difco)  2.0     g
Trypticase peptone (BD-BBL)  2.0     g
Trace vitamins(see Medium No. 197)  10.0     ml
Trace element solution (see Medium No. 243)  10.0     ml
Na2CO3  5.0     g
L-Cysteine·HCl·H2O  0.5     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients, except Na2CO3, L-cysteine·HCl·H2O and Na2S·9H2O. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v) and add Na2CO3. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the L-cysteine·HCl·H2O and Na2S·9H2O solutions. Check pH of the medium to be around 7.5. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20, v/v).