Medium data

Search for medium no. = [431].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

431   BN3 MEDIUM

Basal medium (see below)  1.0     L
Trace element solution (see below)  1.0     ml
Selenite-tungstate solution (see below)  1.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
Vitamin B12  5.0     µg
Glucose  3.6     g
Yeast extract (BD-Difco)  0.5     g
Tryptone (BD-Difco)  1.5     g
Sulfur (powder)  20.0     g
NaHCO3  3.4     g
Resazurin  0.5     mg
Na2S·9H2O  0.36    g
Autoclave Basal medium supplemented with Trace element solution, Selenite-tungstate solution and resazurin under N2-CO2 (4:1, v/v). Separately autoclave glucose, yeast extract and tryptone (10% each) and add to the medium. Filter-sterilize Trace vitamins and NaHCO3 (8%) solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels containing sulfur under a N2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Before inoculation, reduce the medium with 5% Na2S·9H2O solution (autoclaved under a N2 atmosphere). Readjust pH to 7.2, if necessary.
Basal medium:
NaCl  1.0     g
MgCl2·6H2O  0.4     g
CaCl2·2H2O  0.075   g
NH4Cl  0.25    g
KH2PO4  0.2     g
KCl  0.5     g
Distilled water  1.0     L
Trace element solution:
Distilled water  987.0     ml
25% HCl  12.5     ml
FeSO4·7H2O  2.1     g
H3BO3  30.0     mg
MnCl2·4H2O  100.0     mg
CoCl2·6H2O  190.0     mg
NiCl2·6H2O  24.0     mg
CuCl2·2H2O  2.0     mg
ZnSO4·7H2O  144.0     mg
Na2MoO4·2H2O  36.0     mg
Selenite-tungstate solution:
NaOH  0.4     g
Na2SeO3·5H2O  6.0     mg
Na2WO4·2H2O  8.0     mg
Distilled water  1.0     L