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Main sol. 1011c

Internal Solution-ID: S27

Equipment needed: Hungate tubes Autoclave Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 30.00 g 29.412 503.26
2 0.14 g 0.137 0.788
3 0.14 g 0.137 0.934
4 3.40 g 3.333 13.524
5 4.18 g 4.098 20.157
6 0.33 g 0.324 4.34
7 0.25 g 0.245 4.582
8 0.01 g 0.01 0.025
9 Trace element solution 10.00 ml - -
10 1.50 g 1.471 13.875
11 0.50 g 0.49 4.455
12 0.50 g 0.49 4.374
13 0.10 g 0.098 -
14 1.50 g 1.471 5.925
15 Vitamin solution 10.00 ml - -
16 1000.00 ml 978.634 54322.4
17 20.00 mg/l 0.02 -
18 , might need strain adjustment 1.00 g/l - -
Instructions:
  1. Dissolve ingredients (except carbonate, pyruvate, lactate, yeast extract, thiosulfate and vitamins), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials up to a volume of 20% and autoclave. Add pyruvate, lactate, yeast extract, thiosulfate and vitamins to the autoclaved medium 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. Solutions of vitamins and thiosulfate are sterilized by filtration. Adjust pH of the complete medium to 6.7. Prior to inoculation reduce medium with 10 - 20 mg/l sodium dithionite, added from a 5%(w/v) solution freshly prepared under N2 and filter sterilized. After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
1011c DESULFOTHERMUS MJ MEDIUM (H2/CO2)