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).