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Main sol. 120d

Internal Solution-ID: S218

Benötigtes Equipment: Autoclave Hungate tubes Filter Gassing station
Zutat Menge Einheit Konz. [g/L] Konz. [mM]
K2HPO4 0.35 g 0.35 2.009
KH2PO4 0.23 g 0.23 1.69
NH4Cl 0.50 g 0.5 9.348
MgCl2 x 6 H2O 10.00 g 10 49.188
CaCl2 x 2 H2O 0.15 g 0.15 1.02
NaCl 23.00 g 23 393.566
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
2.00 ml 0.002 0.007
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Na2CO3 1.00 g 1 9.435
Wolin's vitamin solution 10.00 ml - -
Methanol 10.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.3 -
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 minto make it anoxic. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N2 gas atmosphere as concentrated solutions in tightly closed tubes. Carbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved separately. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.
120d METHANOSARCINA MARINE MEDIUM