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Main sol. 295

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Internal Solution-ID: S562

Equipment needed: Autoclave Filter Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.199 1.465
NH4Cl 0.25 g 0.249 4.66
NaCl 1.00 g 0.997 17.06
NaNO3 0.80 g 0.798 9.384
MgCl2 x 6 H2O 0.40 g 0.399 1.962
KCl 0.50 g 0.499 6.687
CaCl2 x 2 H2O 0.15 g 0.15 1.017
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.997 9.407
D-Glucose 1.00 g 0.997 5.534
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.703
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, glucose, vitamins, and cysteine, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose, vitamins (sterilized by filtration), and cysteine 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 mixture. Prior to use adjust pH of complete medium to 7.3 - 7.5, if necessary.
2 Note: For transfers use 5 - 10% (v/v) inoculum.
295 OPITUTUS MEDIUM