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Main sol. J976

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Internal Solution-ID: S5000

Equipment needed: Autoclave Anaerobic workbench Filter
1 Mix components thoroughly and adjust pH to 6.75 with NaOH. Distribute the medium into culture vessels (e.g., 5.0 ml in 25 ml serum bottles/Balch tubes) under a stream of N2, seal with butyl rubber stoppers and autoclave. After cooling, aseptically add per liter the following solutions (filter-sterilized and stored anaerobically):
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 3.000 g 2.927 50.083
KCl 0.150 g 0.146 1.963
Na2SO4 0.300 g 0.293 2.061
MgSO4 x 7 H2O 0.123 g 0.12 0.487
CaCl2 x 2 H2O 0.015 g 0.015 0.1
NH4Cl 0.107 g 0.104 1.952
NaH2PO4 1.380 g 1.346 11.222
Mineral solution 5.000 ml - -
Distilled water 1000.000 ml - -
Trace vitamins 10.000 ml - -
Peptone
(w/v, 10%) 		10.000 ml - -
2 Before inoculation, add filter-sterilized O2 gas or air, at a final concentration of 1% O2 in the gas phase.
J976 THERMOFLEXUS HUNGENHOLTZII MEDIUM