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Solution A

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Internal Solution-ID: S4390

Equipment needed: Autoclave Anaerobic workbench Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.3 g 0.333 2.444
K2HPO4 0.3 g 0.333 1.91
NH4Cl 1.0 g 1.109 20.726
Na2SO4 3.0 g 3.326 23.416
NaCl 23.0 g 25.499 436.326
KCl 0.1 g 0.111 1.487
CaCl2 0.1 g 0.111 0.999
Yeast extract
(BD-Difco) 		0.1 g 0.111 -
Trace element solution 1.0 ml - -
L-Cysteine HCl x H2O 0.5 g 0.554 -
Resazurin 1.0 mg 0.001 0.005
Distilled water 900.0 ml - -
1 To complete the medium, add the following stock solutions [Filter-sterilize NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. The other solutions are autoclaved and stored anaerobically under a N2 atmosphere] to the medium (per 4.5 ml):
Sodium lactate
(1 M) 		0.1 ml - -
MgCl2 x 6 H2O
(15%) 		0.1 ml - -
NaHCO3
(5%) 		0.2 ml - -
Na2S x 9 H2O
(2%) 		0.1 ml - -
2 Mix ingredients and adjust pH to 7.2. Distribute 0.9 volume of the medium into tubes (e.g., 4.5 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave at 110C for 45 min.
J565 DESULFOVIBRIO HEB233 MEDIUM