Medium data

Search for medium no. = [807].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

807 WR MEDIUM FOR METHANOBACTERIACEAE

KH₂PO₄	`0.136`	g
NH₄Cl	`0.535`	g
MgCl₂·6H₂O	`0.204`	g
CaCl₂·2H₂O	`0.147`	g
NaHCO₃	`2.52`	g
Tryptone (BD-Difco)	`1.0`	g
Sodium acetate	`0.82`	g
Trace element solution (see Medium No. 243)	`10.0`	ml
Trace vitamins (see Medium No. 197)	`10.0`	ml
Se/W solution (see Medium No. 284)	`1.0`	ml
L-Cysteine·HCl·H₂O	`0.12`	g
Na₂S·9H₂O	`0.12`	g
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix ingredients except NaHCO₃, L-cysteine·HCl·H₂O and Na₂S·9H₂O, and adjust pH to 7.0. Bring to a boil for 5-10 sec, then, cool down under a stream of H₂-CO₂ (80:20, v/v) and add NaHCO₃. Dispense the medium into suitable culture vessels (e.g., 5 ml in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave L-cysteine·HCl·H₂O and Na₂S·9H₂O as 5% solutions, respectively, under a N₂ atmosphere. Prior to inoculation, aseptically and anaerobically add L-cysteine·HCl·H₂O and Na₂S·9H₂O solutions. Readjust pH to 6.7, if necessary. Pressurize the inoculated bottles to 150 kPa H₂-CO₂ (80:20, v/v).