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Main sol. 377b

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Internal Solution-ID: S719

Equipment needed: Hungate tubes Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 13.85 g 13.512 231.215
MgSO4 x 7 H2O 3.50 g 3.415 13.854
MgCl2 x 6 H2O 2.75 g 2.683 13.197
KCl 0.33 g 0.322 4.319
NaBr 0.05 g 0.049 0.474
H3BO3 15.00 mg 0.015 0.237
SrCl2 x 6 H2O
(0.1% w/v) 		7.00 ml 0.007 0.026
(NH4)2SO4 10.00 mg 0.01 0.074
Citric acid
(0.1% w/v) 		5.00 ml 0.005 0.025
KI
(0.01% w/v) 		0.50 ml 4.9e-5 1.9e-4
CaCl2 x 2 H2O 0.75 g 0.732 4.977
KH2PO4 0.50 g 0.488 3.585
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 4.9e-4 0.002
Sulfur
(powdered) 		30.00 g 29.268 912.638
Peptone
(BD Bacto) 		5.00 g 4.878 -
Yeast extract
(OXOID) 		1.00 g 0.976 -
NaHCO3 2.00 g 1.951 23.227
Na2S x 9 H2O 0.50 g 0.488 2.031
Distilled water 1000.00 ml - -
1 Dissolve ingredients except sulfur, peptone, yeast extract, bicarbonate, and sulfide, adjust the pH to 6.8, and sparge medium with 80% N2 and 20% CO2 gas mixture 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, yeast extract, and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust the pH of the complete medium to 6.8, if necessary.
377b PYROCOCCUS ST04 MEDIUM