Medium data

Search for medium no. = [1146].

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

1146 METHANIMICROCOCCUS MEDIUM

NH₄Cl	`0.5`	g
MgSO₄·7H₂O	`0.5`	g
CaCl₂·2H₂O	`0.25`	g
NaCl	`2.25`	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 (Oxoid)	`2.0`	g
Casitone (BD-BBL)	`2.0`	g
Sodium acetate	`1.0`	g
Resazurin	`0.5`	mg
Distilled water	`925.0`	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):

Phosphate solution (see below)	`10.0`	ml
8% NaHCO₃ solution*	`35.0`	ml
50% (v/v) Methanol	`20.0`	ml
1.4% Coenzyme M solution*	`10.0`	ml
Trace vitamins* (see Medium No. 197)	`10.0`	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	`7.2`	ml
5% L-Cysteine·HCl·H₂O solution	`7.2`	ml

Readjust pH to 7.0 - 7.2, if necessary. Pressurize inoculated culture vessels to 100 kPa H₂-CO₂ (80:20, v/v).
Phosphate solution:

K₂HPO₄	`0.35`	g
KH₂PO₄	`0.23`	g
Distilled water	`10.0`	ml