Medium data

Search for medium no. = [1304].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1304   FRESHWATER MEDIUM WITH RHAMNOSE

NaCl  1.0     g
MgCl2·6H2O  0.4     g
CaCl2·2H2O  0.15    g
NH4Cl  0.25    g
KH2PO4  0.2     g
KCl  0.5     g
Resazurin  0.5     mg
Distilled water  955.0     ml
Mix components thoroughly, adjust pH to 7.2-7.4 and autoclave under a N2-CO2 (80:20, v/v) gas mixture. After cooling, while gassing with the same gas mixture, add the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized):
8% NaHCO3 solution*  30.0     ml
Modified trace element mixture (see below)  1.0     ml
Vitamin solution* (see Medium No. 556)  1.0     ml
Selenite-tungstate solution (see Medium No. 431)  0.5     ml
0.5 M L-Rhamnose solution*  10.0     ml
10 % Yeast extract solution  5.0     ml
Readjust pH to 7.2-7.4, if necessary. Aseptically and anaerobically distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. Prior to use, add per liter the following solution (autoclaved and stored under N2):
5% Na2S·9H2O solution  5.0     ml
Modified trace element mixture:
EDTA·Na2  5.2     g
FeCl2·4H2O  1.0     g
H3BO3  30.0     mg
MnCl2·4H2O  100.0     mg
CoCl2-6H2O  190.0     mg
NiCl2-6H2O  24.0     mg
CuCl2-2H2O  2.0     mg
ZnSO4-7H2O  144.0     mg
Na2MoO4-2H2O  36.0     mg
Distilled water  1.0     L
Dissolve EDTA·Na2 in distilled water first and adjust pH to 6.5 with NaOH. Then, proceed to add minerals.



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