Medium data

Search for medium no. = [994].

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

994 MBM MEDIUM 2

NaCl	`80.0`	g
MgSO₄·7H₂O	`6.0`	g
KCl	`1.5`	g
NH₄Cl	`1.0`	g
CaCl₂·2H₂O	`0.4`	g
K₂HPO₄	`0.4`	g
Trace minerals (see Medium No. 151)	`10.0`	ml
Sodium acetate·3H₂O	`1.6`	g
Resazurin	`1.0`	mg
Distilled water	`900.0`	ml

Mix components thoroughly. Bring to a boil for a few seconds, cool down under a N₂-CO₂ (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add the following solutions (autoclaved or *filter-sterilized):

1 M Disodium fumarate solution	`40.0`	ml
8% NaHCO₃ solution*	`50.0`	ml
Trace vitamins* (see Medium No. 197)	`10.0`	ml

Aseptically distribute the medium into culture vessels (e.g., 5 ml of the medium in the Hungate tubes) under a N₂-CO₂ (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N₂ gas atmosphere):

5% L-Cysteine·HCl·H₂O solution	`6.0`	ml
5% Na₂S·9H₂O solution	`6.0`	ml

Readjust pH to 7.5, if necessary.