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Main sol. 329

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Internal Solution-ID: S605

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
K2SO4 0.01 g 0.01 0.057
NaCl 70.00 g 69.86 1195.42
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID) 		0.05 g 0.05 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 5.00 g 4.99 59.4
MgCl2 x 6 H2O 0.33 g 0.329 1.62
CaCl2 x 2 H2O 0.33 g 0.329 2.24
Trimethylamine-HCl 5.00 g 4.99 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.705
Na2S x 9 H2O 0.15 g 0.15 0.623
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, magnesium chloride, calcium chloride, vitamins, trimethylamine, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and equilibrate pH to 6.9 - 7.0, then distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add magnesium chloride, calcium chloride, vitamins (sterilized by filtration), trimethylamine, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use adjust pH of complete medium to 7.2, if necessary.
329 METHANOHALOPHILUS HALOPHILUS MEDIUM