Medium data

Search for medium no. = [420].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

420   MJAIS/MD MEDIUM

NaCl  30.0     g
K2HPO4  0.09    g
KH2PO4  0.07    g
CaCl2·2H2O  0.8     g
NH4Cl  1.25    g
MgSO4·7H2O  3.4     g
MgCl2·6H2O  4.18    g
KCl  0.33    g
NiCl2·6H2O  10.0     mg
Na2SeO3·5H2O  10.0     mg
Na2WO4·2H2O  10.0     mg
Fe(NH4)2(SO4)2·6H2O  10.0     mg
FeSO4·7H2O  20.0     mg
Trace mineral solution (see below)  10.0     ml
Trace vitamins (see Medium No. 197)  1.0     ml
NaHCO3  2.0     g
Sulfur (powder)  30.0     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients, except Trace vitamins, NaHCO3, sulfur and Na2S·9H2O, adjust pH to 6.0, and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution, and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S·9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml of medium in 120 ml serum bottles containing 0.6 g sulfur) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the sterile Na2S·9H2O solution. Pressurize the inoculated bottle to 300 kPa H2-CO2 (4:1, v/v).
Trace mineral solution:
Use Trace mineral solution of Medium No. 324 supplemented with 0.01 g/L SrCl2·6H2O, 0.01 g/L NaBr and 0.01 g/L KI.