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Main sol. 1043

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Internal Solution-ID: S2111

Equipment needed: Autoclave Serum bottle Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.286 9.731
CaCl2 x 2 H2O 74.00 mg 0.073 0.498
MgSO4 x 7 H2O 0.28 g 0.277 1.124
KH2PO4 0.28 g 0.277 2.035
Yeast extract
(OXOID) 		0.10 g 0.099 -
Trypticase peptone
(BD BBL) 		2.00 g 1.978 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
FeCl3 x 6 H2O
(0.1% w/v in 0.2 N HCl) 		0.50 ml 4.9e-4 0.002
L-Cysteine HCl x H2O 0.30 g 0.297 1.689
Starch
(soluble) 		2.00 g 1.978 -
Na2S x 9 H2O 0.30 g 0.297 1.235
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except ferric chloride, cysteine, starch and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add ferric chloride and solid cysteine, then adjust pH to 6.3. Dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 50 ml bottles) and autoclave. Prior to inoculation add starch and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 6.5, if necessary.
1043 IGNISPHAERA MEDIUM