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Main sol. 319

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Internal Solution-ID: S593

Equipment needed: Hungate tubes Autoclave Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Casamino acids
(BD Bacto) 		2.00 g 1.974 -
Nutrient broth
(BD Difco) 		2.00 g 1.974 -
Yeast extract 2.00 g 1.974 -
L-Glutamic acid 4.00 g 3.949 26.838
NaCl 105.00 g 103.653 1773.66
KCl 0.75 g 0.74 9.931
Modified Wolin's mineral solution 10.00 ml - -
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.00 ml 0.002 0.007
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
MgCl2 x 6 H2O 10.00 g 9.872 48.556
CaCl2 x 2 H2O 3.70 g 3.653 24.845
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except cysteine, magnesium chloride, calcium chloride and vitamins), bring medium to the boil, then cool to room temperature under 100% N2 gas atmosphere. Thereafter, add and dissolve cysteine, adjust pH to 6.5 and dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add magnesium chloride and calcium chloride from sterile anoxic stock solutions autoclaved under 100% N2 gas and vitamins from a filter-sterilized stock solution prepared under N2 gas.
2 Note: For agar medium add 20.00 g/l agar, 5.00 g/l CaCO3, and 2.00 g/l soluble starch.
319 SPOROHALOBACTER LORTETII MEDIUM