Medium data

Search for medium no. = [1408].

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

1408 TMBS4 MEDIUM

NaCl	`1.0`	g
MgCl₂·6H₂O	`0.4`	g
CaCl₂·2H₂O	`0.15`	g
KCl	`0.5`	g
NH₄Cl	`0.25`	g
KH₂PO₄	`0.2`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Selenite tungstate solution (see Medium No. 431)	`1.0`	ml
Resazurin	`0.5`	mg
Distilled water	`930.0`	ml

Mix components thoroughly and autoclave under a N₂-CO₂ (4:1, v/v) gas atmosphere. After cooling, add the following solutions from anaerobic stocks in the sequence as indicated (autoclaved or *filter-sterilized):

5% NaHCO₃ solution*	`30.0`	ml
Trace vitamins* (see Medium No. 197)	`10.0`	ml
Vitamin B₁₂ solution* (see Medium No. 403)	`1.0`	ml
5% Sodium thiosulfate solution	`2.0`	ml
10% Yeast extract solution	`10.0`	ml
Syringate solution (see below)	`10.0`	ml
0.1 M Dithiothreitol solutuion*	`10.0`	ml

Aseptically and anaerobically, distribute the medium into culture vessels under the same gas mixture (e.g., 20 ml in 60 ml-serum bottles) and seal with butyl rubber stoppers. The final pH of the medium should be 7.2-7.4. After inoculation, add per liter the following solution (filter-sterilized, freshly prepared):

2.5% Sodium dithionite solution 10.0 ml

Syringate solution:
Dissolve 6 g syringic acid in 70 ml distilled water and adjust pH to 7.5 with 2 N NaOH solution. Fill up to 100 ml with distilled water.