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Main sol. 498

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Internal Solution-ID: S1025

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract
(BD Bacto) 		0.50 g 0.488 -
Modified Wolin's mineral solution 15.00 ml - -
NaCl 3.46 g 3.379 57.818
Na2-EDTA 0.77 g 0.752 2.237
MgSO4 x 7 H2O 0.88 g 0.859 3.487
MgCl2 x 6 H2O 0.69 g 0.674 3.314
CaCl2 x 2 H2O 0.14 g 0.137 0.93
KCl 0.08 g 0.078 1.048
NaBr 12.50 mg 0.012 0.119
H3BO3
(0.1% w/v) 		4.00 ml 0.004 0.063
(NH4)2Ni(SO4)2
(0.1% w/v) 		3.00 ml 0.003 0.007
SrCl2 x 6 H2O
(0.1% w/v) 		1.90 ml 0.002 0.007
KI
(0.01% w/v) 		0.10 ml 9.8e-6 3.8e-5
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
KH2PO4 0.50 g 0.488 3.588
Starch
(soluble) 		5.00 g 4.883 -
Na2S x 9 H2O 0.50 g 0.488 2.033
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except hydrogenphosphate, starch and sulfide) and adjust pH to 7.0. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% volume and autoclave. Add hydrogenphosphate, starch and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Check pH of the complete medium and adjust to 7.0 - 7.2, if necessary.
498 PSEUDOTHERMOTOGA MEDIUM