2
Mix solution D, C and E in the cotton-stoppered Erlenmeyer flask. Bubble with CO2 in an ice bath under sterile conditions (5-10 min).
3
Fill 50 ml of the mixed solution (C,D,E) into each 100 ml bottle of solution A (for a total volume of approx. 96 ml).
4
Before using, add 2,6 ml solution B (Na2S x 9 H2O), 0.1 ml solution F (Vitamin B12) and 1 ml of solution G (2,5% Ammonium-acetate and Magnesium-acetate) to each 100ml bottle.
5
Adjust the pH with filter-sterilised 1M Na2CO3 to 7.1-7.3.
6
Fill in sterile, N2 gassed screw-cap tubes under N2 gas.
7
For the growth of DSM 11081 feed the culture with neutralized sulfide feeding solution regularly (approx. every 3 days). 0,5-1 ml/ 100 mL
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