Medium data

Search for medium no. = [1147].

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

1147 METHANOBACTERUM MEDIUM (IX)

KH₂PO₄	`0.5`	g
MgSO₄·7H₂O	`0.4`	g
NaCl	`0.4`	g
NH₄Cl	`0.4`	g
CaCl₂·2H₂O	`0.05`	g
FeSO₄·7H₂O	`0.002`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Yeast extract	`1.0`	g
Sodium acetate	`1.0`	g
Sodium formate	`2.0`	g
Fatty acid mixture (see Medium No. 266)	`20.0`	ml
Resazurin	`0.5`	mg
Distilled water	`922.0`	ml

Mix components and autoclave under a N₂-CO₂ (80:20, v/v) gas atmosphere. After cooling, add the following solutions from anaerobic stocks (autoclaved or filter-sterilized):

Selenite-tungstate solution* (see Medium No. 431)	`0.5`	ml
8% NaHCO₃ solution*	`50.0`	ml
50% (v/v) Methanol	`6.3`	ml

Aseptically and anaerobically distribute the medium into culture vessels under a H₂-CO₂ (80:20, v/v) gas stream and seal with butyl stoppers. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N₂ atmosphere):

5% Na₂S·9H₂O solution	`10.0`	ml
5% L-Cysteine·HCl·H₂O solution	`10.0`	ml

Readjust pH to 7.5, if necessary. Pressurize inoculated culture vessels to 100 kPa H₂-CO₂ (80:20, v/v).