KH2PO4 0.20g NH4Cl 0.25g NaCl 20.00g MgCl2 x 6 H2O 3.00g KCl 0.50g CaCl2 x 2 H2O 0.15g Trace element solution SL-10 1.00ml Sodium resazurin (0.1% w/v) 0.50ml Na2CO3 1.25g Disodium succinate 3.25g Na2S x 9 H2O 0.36g Distilled water 1000.00ml
1. Dissolve ingredients except carbonate, succinate and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Add succinate (sterilized by filtration) and sulfide 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. Prior to use adjust pH of complete medium to 7.2 - 7.5.
2. Note: Dispense medium into anoxic Hungate-type tubes or serum vials only to 30% of their volume to allow a large head space.
For DSM 2376: Supplement medium with 0.10% yeast extract (separately sterilized). For DSM 2377, DSM 2378: Replace Na2-succinate with 0.68 g/l of gallic acid. For DSM 2380: Replace Na2-succinate with 0.90 g/l of 2,3-butanediol. For DSM 2382: Replace Na2-succinate with 2.00 g/l of sodium L-tartrate. For DSM 2394: Replace Na2-succinate with 1.00 g/l of polyethylene glycol (molecular weight 106 - 20000). For DSM 3247: Replace Na2-succinate with 1.00 g/l acetoin. For DSM 6831: Supplement medium with 1.00 ml/l of a Wolin's vitamin solution 10x (see medium 120) and replace Na2-succinate with 1.00 g/l of quinic acid added from a neutralized anoxic stock solution (0.5 M). For DSM 105538: Replace Na2-succinate with 2.00 g/l of D-glucose and supplement medium with 1.00 g/l yeast extract and 1.00 ml/l of a Wolin's vitamin solution 10x (see medium 120) added from sterile anoxic stock solutions.
Trace element solution SL-10 (from medium 320) HCl (25%) 10.00ml FeCl2 x 4 H2O 1.50g ZnCl2 70.00mg MnCl2 x 4 H2O 100.00mg H3BO3 6.00mg CoCl2 x 6 H2O 190.00mg CuCl2 x 2 H2O 2.00mg NiCl2 x 6 H2O 24.00mg Na2MoO4 x 2 H2O 36.00mg Distilled water 990.00ml First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
