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Main sol. J1248

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Internal Solution-ID: S5384

Equipment needed: Anaerobic workbench Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.000 g 0.944 16.158
MgCl2 x 6 H2O 0.500 g 0.472 2.322
KH2PO4 0.200 g 0.189 1.388
NH4Cl 0.300 g 0.283 5.296
KCl 0.300 g 0.283 3.8
CaCl2 x 2 H2O 0.015 g 0.014 0.096
FeCl2 solution 1.000 ml - -
Trace element solution 1.000 ml - -
Selenite-tungstate solution 1.000 ml - -
Sodium formate 0.340 g 0.321 4.721
Sodium acetate 0.410 g 0.387 4.72
Resazurin 0.250 mg 2.4e-4 0.001
Distilled water 1000.000 ml - -
1 Mix components thoroughly, bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas mixture. Add the following solutions:
NaHCO3
(8%) 		30.000 ml - -
L-Cysteine HCl x H2O
(5%) 		6.000 ml - -
Na2S x 9 H2O
(5%) 		10.000 ml - -
2 Adjust pH of the medium to 7.0-7.2, if necessary. Dispense the medium into culture vessels (e.g., 100 ml in 120 ml-serum bottles) under the same gas mixture and seal with butyl rubber stoppers or Teflon-lined stoppers. Then, anaerobically add per liter the following solution:
Tetrachloroethene 60.000 μl - -
3 Finally, add per liter the following solution from an anaerobic stock (filter-sterilized):
Trace vitamins 10.000 ml - -
4 Dissolve tetrachloroethene with ultrasound. Autoclave in a closed basket. Do not remove the culture vessels from autoclave before temperature of the media has reached below 70C.
J1248 MINERAL SALTS MEDIUM WITH TCE