Medium data

Search for medium no. = [230].

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

230 METHANOSARCINA MEDIUM

KH₂PO₄	`0.227`	g
K₂HPO₄	`0.348`	g
NH₄Cl	`0.5`	g
MgSO₄·7H₂O	`0.5`	g
CaCl₂·2H₂O	`0.25`	g
NaCl	`2.25`	g
FeSO₄·7H₂O	`2.0`	mg
Yeast extract (BD-Difco)	`2.0`	g
Casitone (BD-Difco)	`2.0`	g
Trace vitamins (see Medium No. 197)	`10.0`	ml
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
NaHCO₃	`0.85`	g
Methanol	`10.0`	ml
L-Cysteine·HCl·H₂O	`0.3`	g
Na₂S·9H₂O	`0.3`	g
Resazurin	`1.0`	mg
Distilled water	`980.0`	ml

Mix ingredients, except NaHCO₃, methanol, cysteine·HCl and Na₂S·9H₂O, and bring to a boil for 5-10 sec. Cool down under a stream of N₂-CO₂ (80:20), and add NaHCO₃ and methanol to the medium. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of N₂-CO₂ (80:20) 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 cysteine·HCl and Na₂S·9H₂O solutions.