Medium data

Search for medium no. = [244].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

244   STETTERIA MEDIUM

Synthetic seawater (2 x, see below)  500.0     ml
NaHCO3  0.16    g
KH2PO4  0.5     g
Trace minerals (see Medium No. 151)  15.0     ml
Ni-Se-W solution (see below)  10.0     ml
Bacto peptone (BD-Difco)  2.0     g
Yeast extract (BD-Difco)  1.0     g
Resazurin  0.75    mg
Na2S·9H2O  0.5     g
Sulfur (powder)  10.0     g
Distilled water  500.0     ml
Mix ingredients, except sulfur and Na2S·9H2O, and adjust pH to 6.0 with H2SO4. Sterilize the medium by filtration. Steam sulfur for 3 hr on each of 3 successive days. Neutralize Na2S·9H2O as a 5% solution, and autoclave under a N2 atmosphere. Distribute the medium and sulfur into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S·9H2O solution, and readjust pH to 6.0 with sterile 1 N H2SO4. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).
Synthetic seawater (2 x):
NaCl  55.4     g
MgSO4·7H2O  14.0     g
MgCl2·6H2O  11.0     g
KCl  1.3     g
NaBr  0.2     g
H3BO3  0.06    g
SrCl2·6H2O  0.03    g
KI  0.1     mg
CaCl2·2H2O  1.5     g
Distilled water  1.0     L
Ni-Se-W solution:
NiCl2·6H2O  25.0     mg
(NH4)2Ni(SO4)2·6H2O  2.0     g
Na2SeO3  0.3     mg
Na2SeO4  10.0     mg
Na2WO4·2H2O  10.0     mg
Distilled water  1.0     L


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