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Main sol. J417

Internal Solution-ID: S4151

Equipment needed: Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 20.00 g 19.782 338.508
K2HPO4 0.90 g 0.89 5.111
KH2PO4 0.70 g 0.692 5.088
CaCl2 x 2 H2O 1.00 g 0.989 6.728
NH4Cl 1.25 g 1.236 23.115
MgSO4 x 7 H2O 3.40 g 3.363 13.644
MgCl2 x 6 H2O 4.18 g 4.135 20.337
KCl 0.33 g 0.326 4.378
NiCl2 x 6 H2O 5.00 mg 0.005 0.038
Na2SeO3 x 5 H2O 5.00 mg 0.005 0.019
Fe2(SO4)3 x n H2O 0.01 g 0.01 -
H2WO4 5.00 mg 0.005 0.02
FeSO4 x 7 H2O 0.02 g 0.02 0.071
Yeast extract
(BD-Difco)
1.00 g 0.989 -
Tryptone
(BD-Difco)
1.00 g 0.989 13.916
Sodium formate 0.20 g 0.198 2.909
Trace mineral solution 10.00 ml - -
Trace vitamins 1.00 ml - -
NaHCO3 2.00 g 1.978 23.549
Sulfur
(powder)
10.00 g 9.891 308.425
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Mix ingredients, except Trace vitamins, NaHCO3 and sulfur, 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 x 9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) containing sterilized sulfur under a stream of H2-CO2 (4:1, v/v) gas mixture, sparge for 5 min with the same gas mixture, and seal with butyl rubber stoppers. Prior to inoculation, add the sterilized Na2S x 9H2O solution to the medium, and pressurize to 200 kPa H2-CO2 (4:1, v/v). Readjust pH to 5.5, if necessary.
J417 MJYTFSH MEDIUM