Medium data

Search for medium no. = [470].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

470   VULCANIBACILLUS MEDIUM

Sea salts (Sigma)  30.0     g
NaNO3  1.0     g
Wolfe's mineral elixir (see below)  2.0     ml
VOSO4·xH2O  0.05    mg
Sodium pyruvate  1.0     g
Resazurin  0.5     mg
FeSO4·7H2O  0.1     g
NaHCO3  1.0     g
Yeast extract  0.5     g
Trace vitamins (see Medium No. 197)  20.0     ml
Distilled water  1.0     L
Dissolve ingredients except FeSO4·7H2O, NaHCO3, yeast extract and vitamins, and adjust pH to 7.0. Bring to a boil for 3 min, then cool to room temperature under a N2 gas stream. Dispense the medium under the same gas atmosphere into culture vessels, seal with butyl rubber stoppers and autoclave. Add sterile anaerobic stock solutions of FeSO4·7H2O (1%, in 0.1 N H2SO4), NaHCO3 (8%, under a N2·CO2 gas mixture), yeast extract (10%) and vitamins (filter-sterilized) to the medium. Adjust final pH of the medium to 7.0 - 7.2. Add 10 - 20 mg sodium dithionite per liter [e.g., from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized], if the medium is not completely reduced after inoculation.
Wolfe's mineral elixir:
MgSO4·7H2O  30.0     g
MnSO4·xH2O  5.0     g
NaCl  10.0     g
FeSO4·7H2O  1.0     g
CoCl2·6H2O  1.8     g
CaCl2·2H2O  1.0     g
ZnSO4·7H2O  1.8     g
CuSO4·5H2O  0.1     g
KAl(SO4)2·12H2O  0.18    g
H3BO3  0.1     g
Na2MoO4·2H2O  0.1     g
(NH4)2Ni(SO4)2·6H2O  2.8     g
Na2WO4·2H2O  0.1     g
Na2SeO4  0.1     g
Distilled water  1.0     L
First adjust pH to 1.0 with diluted H2SO4, then dissolve the salts.



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