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Main sol. 986

Internal Solution-ID: S2015

Equipment needed: Autoclave Filter Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA)
30.00 g 29.97 -
NH4Cl 1.00 g 0.999 18.676
K2HPO4 0.30 g 0.3 1.721
KH2PO4 0.30 g 0.3 2.202
Peptone 0.50 g 0.5 -
Yeast extract 0.50 g 0.5 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
NaHCO3 2.00 g 1.998 23.784
D-Glucose 4.00 g 3.996 22.18
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.25 1.422
Na2S x 9 H2O 0.25 g 0.25 1.04
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, glucose, vitamins, cysteine and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, glucose, sulfide and cysteine 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. Vitamins should be sterilized by filtration. The pH of the complete medium should be 7.0.
986 PARAMALEDIVIBACTER MEDIUM