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Main sol. 1003

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Internal Solution-ID: S2046

Equipment needed: Autoclave Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
Water 1000.00 ml - -
1 Place 1000 ml of anaerobic water (CO2 water) in a flask. Add the following compounds:
MgSO4 x 7 H2O 7.00 g 7 28.4
NaS2O3 2.00 g 2 -
CaCl2 x 2 H2O 0.40 g 0.4 2.721
KCl 0.48 g 0.48 6.439
MgCl2 0.78 g 0.78 8.192
MES 1.95 g 1.95 9.988
2 Add solutions A, B, and Trace mineral solution (see recipes below):
Solution A (100x solution) 2.00 ml - -
Solution B (500x solution) 1.50 ml - -
Trace mineral solution (100x) 10.00 ml - -
3 pH the media to 6.0 and gas the flask containing the media with CO2 for 20 min at least. Gas first culture tube in rack and place 5 ml of media in previous tube. Place stopper on loaded culture tube, crimp tube cap onto stopper. Autoclave the medium for 20 min., 121°C. Add 1 ml of O2 to each tube before inoculation. After the inoculation the tubes are pressurized with H2 (138 KP).
1003 HYDROGEN-OXYDIZING MEDIUM