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Main sol. 718

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Internal Solution-ID: S1469

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.336 4.502
MgCl2 x 6 H2O 4.00 g 3.949 19.423
MgSO4 x 7 H2O 3.45 g 3.406 13.818
NH4Cl 0.25 g 0.247 4.614
CaCl2 x 2 H2O 0.14 g 0.138 0.94
K2HPO4 0.14 g 0.138 0.793
NaCl 18.00 g 17.769 304.055
Modified Wolin's mineral solution 10.00 ml - -
Fe(NH4)2(SO4)2 x 7 H2O
(0.1% w/v) 		2.00 ml 0.002 0.005
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.00 g 0.987 11.751
D-Glucose 5.00 g 4.936 27.397
Yeast extract 0.20 g 0.197 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.494 2.055
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, glucose, yeast extract, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate, adjust pH to 6.5 and dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add glucose, yeast extract, vitamins, and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. Prior to inoculation adjust pH of complete medium to 6.5 - 6.7, if necessary.
718 PETROTOGA MEDIUM