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Main sol. J872

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Internal Solution-ID: S4858

Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.489 3.595
MgSO4 x 7 H2O 0.40 g 0.391 1.588
NaCl 5.00 g 4.892 83.716
NH4Cl 0.40 g 0.391 7.317
CaCl2 x 2 H2O 0.05 g 0.049 0.333
FeSO4 x 7 H2O 2.00 mg 0.002 0.007
FeCl2 solution 1.00 ml - -
Trace element solution 1.00 ml - -
Yeast extract 1.00 g 0.978 -
Sludge fluid 50.00 ml - -
Sodium acetate 1.00 g 0.978 11.928
Sodium formate 2.00 g 1.957 28.776
NaHCO3 4.00 g 3.914 46.59
Fatty acid mixture 20.00 ml - -
Resazurin 1.00 mg 9.8e-4 0.004
Distilled water 930.00 ml - -
1 Mix components except NaHCO3 and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3 to the medium. Dispense the medium into culture vessels under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N2 atmosphere):
Na2S x 9 H2O
(5%) 		10.00 ml - -
L-Cysteine HCl x H2O
(5%) 		10.00 ml - -
2 Pressurize inoculated vessels to 200 kPa with a H2-CO2 (80:20, v/v) gas mixture.
J872 METHANOBACTERIUM MEDIUM (IV)