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Main sol. 318b

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Internal Solution-ID: S591

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.30 g 0.296 2.176
NaCl 0.60 g 0.592 10.135
MgCl2 x 6 H2O 0.10 g 0.099 0.486
CaCl2 x 2 H2O 0.08 g 0.079 0.537
Trace element solution 10.00 ml - -
NH4Cl 1.00 g 0.987 18.455
Yeast extract
(OXOID) 		0.50 g 0.494 -
Trypticase peptone
(BD BBL) 		0.50 g 0.494 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
KHCO3 4.00 g 3.949 39.441
Ethanol 1.30 ml - -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.296 1.686
Na2S x 9 H2O 0.30 g 0.296 1.233
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, ethanol, vitamins, cysteine and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate and adjust pH to 7.0, then dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add ethanol, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins are prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0 - 7.2, if necessary.
318b ANAEROTIGNUM NEOPROPIONICUM MEDIUM