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Main sol. 377

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Internal Solution-ID: S717

Equipment needed: Hungate tubes Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 13.85 g 13.85 236.995
MgSO4 x 7 H2O 3.50 g 3.5 14.2
MgCl2 x 6 H2O 2.75 g 2.75 13.527
KCl 0.33 g 0.33 4.427
NaBr 0.05 g 0.05 0.486
H3BO3 15.00 mg 0.015 0.243
SrCl2 x 6 H2O
(0.1% w/v) 		7.00 ml 0.007 0.026
(NH4)2SO4 10.00 mg 0.01 0.076
Citric acid
(0.1% w/v) 		5.00 ml 0.005 0.026
KI
(0.01% w/v) 		0.50 ml 5.0e-5 1.9e-4
CaCl2 x 2 H2O 0.75 g 0.75 5.102
KH2PO4 0.50 g 0.5 3.674
NiCl2 x 6 H2O
(0.1% w/v) 		2.00 ml 0.002 0.015
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Sulfur
(powdered) 		30.00 g 30 935.454
Peptone
(BD Bacto) 		5.00 g 5 -
Yeast extract
(OXOID) 		1.00 g 1 -
Neutralized sulfide solution 3% (w/v) 15.00 ml - -
Distilled water 960.00 ml - -
1 Dissolve ingredients except for sulfur, peptone, yeast extract, and sulfide, adjust the pH to 6.5 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials that already contain the appropriate amount of sulfur. Sterilize the medium by autoclaving at 110°C for 20 min. After sterilization, add peptone and yeast extract from sterile stock solutions prepared under 100% N2 gas atmosphere and sulfide from a neutralized stock solution.
377 PYROCOCCUS MEDIUM