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Main sol. J1083

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Internal Solution-ID: S5151

Equipment needed: Anaerobic workbench Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2HPO4 x 2 H2O 0.53 g 0.519 3.657
KH2PO4 0.41 g 0.402 2.951
NH4Cl 0.30 g 0.294 5.493
NaCl 0.30 g 0.294 5.028
MgCl2 x 6 H2O 0.10 g 0.098 0.482
CaCl2 x 2 H2O 0.11 g 0.108 0.733
Acid trace element solution 1.00 ml - -
Alkaline trace element solution 1.00 ml - -
Sodium acetate x 3 H2O 0.68 g 0.666 -
Yeast extract 0.10 g 0.098 -
Sulfur
(powder) 		0.80 g 0.784 24.432
Resazurin 1.00 m - -
Distilled water 1000.00 ml - -
1 Mix components, except sulfur, and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized):
Vitamin solution 0.20 ml - -
NaHCO3
(8%) 		12.50 ml - -
2 Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically distribute the medium and sulfur into culture vessels (e.g., 50 ml of the medium and 0.04 g of sulfur in 120 ml serum bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, anaerobically add per liter the following solution from an anaerobic stock (autoclaved and stored under a N2 atmosphere):
Na2S x 9 H2O
(5%) 		6.00 ml - -
3 Adjust pH to 6.0, if necessary.
J1083 DESULFURELLA TR1 MEDIUM