Medium data

Search for medium no. = [368].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

368   MJAIS MEDIUM

NaCl  30.0     g
K2HPO4  0.9     g
KH2PO4  0.7     g
CaCl2·2H2O  1.0     g
NH4Cl  1.25    g
MgSO4·7H2O  3.4     g
MgCl2·6H2O  4.18    g
KCl  0.33    g
NiCl2·6H2O  5.0     mg
Na2SeO3·5H2O  0.5     mg
Fe2(SO4)3·xH2O  0.01    mg
H2WO4  5.0     mg
FeSO4·7H2O  0.02    g
Ferric citrate  0.02    g
Trace mineral solution (see below)  10.0     ml
Trace vitamins (see Medium No. 197)  1.0     ml
NaHCO3  1.0     g
Sulfur (powder)  10.0     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 5.5, 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 200 kPa H2-CO2 (4:1, v/v).
Trace mineral solution:
Nitrilotriacetic acid  1.5     g
MnSO4·xH2O  0.5     g
CoSO4·7H2O  0.5     g
ZnSO4·7H2O  0.18    g
CuSO4·5H2O  0.01    g
AlK(SO4)2  0.02    g
H3BO3  0.01    g
Na2MoO4·2H2O  0.01    g
SrCl2·6H2O  0.01    g
NaBr  0.01    g
KI  0.01    g
Distilled water  1.0     L
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.



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