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Main sol. 643

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Internal Solution-ID: S1367

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sodium Potassium phosphate buffer
(0.2 M, pH 7.0) 		100.00 ml - 20
NaCl 10.00 g 9.94 170.095
NH4Cl 0.50 g 0.497 9.292
MgCl2 x 7 H2O 0.10 g 0.099 1.044
CaCl2 x 2 H2O 0.03 g 0.03 0.203
Modified Wolin's mineral solution 5.00 ml - -
Yeast extract 10.00 g 9.94 -
Glycine 0.75 g 0.746 9.931
L-Arginine x HCl 2.10 g 2.087 -
L-Lysine HCl 1.80 g 1.789 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.40 g 0.398 2.264
Na2S x 9 H2O 0.40 g 0.398 1.655
Distilled water 900.00 ml - -
1 Dissolve ingredients except vitamins, cysteine and sulfide, sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Check pH of complete medium and adjust to pH 7.0, if necessary.
643 SEDIMENTIBACTER MEDIUM