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Main sol. 895

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Internal Solution-ID: S1845

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.00 g 0.987 7.471
NH4Cl 0.40 g 0.395 7.382
Na2S2O3 x 5 H2O 0.10 g 0.099 0.398
K2HPO4 0.50 g 0.494 2.834
MgSO4 x 7 H2O 0.10 g 0.099 0.401
CaCl2 x 2 H2O 0.05 g 0.049 0.336
NaCl 10.00 g 9.872 168.92
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.00 ml 0.002 0.007
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract
(BD Bacto) 		0.25 g 0.247 -
Tryptone
(BD Bacto) 		0.25 g 0.247 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
L-Cysteine HCl x H2O 0.50 g 0.494 2.81
Na2CO3 3.00 g 2.962 27.942
Starch
(soluble) 		2.00 g 1.974 -
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve all ingredients except cysteine, carbonate, starch and vitamins. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, add cysteine and adjust pH to 7.5, then dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add starch and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 9.3 - 9.5, if necessary.
895 ANAEROBRANCA GOTTSCHALKII MEDIUM