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Main sol. 318a

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Internal Solution-ID: S590

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
HEPES
(SIGMA) 		11.92 g 11.79 49.477
KH2PO4 0.30 g 0.297 2.181
NaCl 0.60 g 0.593 10.155
MgCl2 x 6 H2O 0.10 g 0.099 0.487
CaCl2 x 2 H2O 0.08 g 0.079 0.538
Trace element solution 10.00 ml - -
NH4Cl 1.00 g 0.989 18.492
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
KHCO3 2.00 g 1.978 19.76
2-Furoic acid
(SIGMA) 		2.24 g 2.216 19.768
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.297 1.689
Na2S x 9 H2O 0.30 g 0.297 1.235
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, vitamins, furoic acid, cysteine and sulfide and adjust pH of solution to 7.0. 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 furoic acid (stock solution neutralized with NaOH), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.0 - 7.2, if necessary.
318a PELOBACTER (FUF) MEDIUM