Medium data

Search for medium no. = [1427].

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

1427 MODIFIED METHANOBACTERIUM MEDIUM

KH₂PO₄	`0.5`	g
MgSO₄·7H₂O	`0.4`	g
NaCl	`0.4`	g
NH₄Cl	`0.4`	g
CaCl₂·2H₂O	`0.05`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Brain heart infusion (BD Difico)	`6.0`	g
Proteose peptone (BD Difico)	`6.0`	g
Yeast extract (Oxoid)	`2.0`	g
Sodium acetate	`1.0`	g
Sodium formate	`2.0`	g
Rumen fluid, clarified (see Medium No. 266)	`10.0`	ml
Resazurin	`0.5`	mg
Distilled water	`940.0`	ml

Mix components and autoclave under a N₂-CO₂ (80:20, v/v) gas mixture. After cooling, aseptically and anaerobically, add the following solutions from anaerobic stocks (filtersterilized):

Vitamin solution (see Medium No. 898)	`1.0`	ml
8% NaHCO₃ solution	`50.0`	ml

Dispense the medium into culture vessels (e.g., 5 ml medium in Hungate tubes) 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

After inoculation, pressurize the inoculated culture vessels to 100 kPa H₂-CO₂ (80:20, v/v).