Medium data

Search for medium no. = [375].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

375   MARINE MEDIUM WITH SULFUR

NaCl  30.0     g
MgCl2·6H2O  3.0     g
KCl  0.5     g
NH4Cl  0.5     g
NaNO3  1.0     g
KH2PO4  0.2     g
MES  1.95    g
CaCl2·2H2O  0.15    g
Sulfur  5.0     g
Na2S·9H2O  0.5     g
Distilled water  1.0     L
Mix ingredients, except sulfur and Na2S·9H2O, adjust pH to 6.0 with NaOH, and autoclave. Steam sulfur for 3 hr on each of 3 successive days. Neutralize 5% Na2S·9H2O solution and autoclave under a N2 atmosphere. Distribute the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) containing sulfur 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 solution to the medium, and readjust pH, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).



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