Medium data

Search for medium no. = [1125].

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

1125 ARTFICIAL FRESHWATER MEDIUM II

KH₂PO₄	`0.2`	g
NH₄Cl	`0.25`	g
KCl	`0.3`	g
Na₂SO₄	`1.4`	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	`1.0`	mg
Distilled water	`960.0`	ml

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

8% NaHCO₃ solution*	`30.0`	ml
1 M MgCl₂·6H₂O solution	`2.5`	ml
1 M CaCl₂·2H₂O solution	`1.0`	ml
Vitamin solution (see Medium No. 403)*	`1.0`	ml
Thiamine solution (see Medium No. 403)*	`1.0`	ml
Vitamin B₁₂ solution (see Medium No. 403)*	`1.0`	ml
10% Yeast extract solution	`1.0`	ml
1 M Glycerin solution*	`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. Prior to use, add per liter the following solution to reduce the medium (autoclaved and stored under N₂):

5% Na₂S·9H₂O solution 5.0 ml

Comment:
For cultivation of JCM 31104, use 1 M sodium lactate solution instead of 1 M glycerin solution (final 10 mM).