培養(yǎng)基配方
Final pH: 7.0 - 7.2
Final volume: 1003 ml
NaCl 1.17g
MgCl2 x 6 H2O 0.40g
KCl 0.30g
CaCl2 x 2 H2O 0.15g
NH4Cl 0.27g
KH2PO4 0.20g
Na2SO4 2.84g
Trace element solution SL-10 1.00ml
Selenite-tungstate solution 1.00ml
Yeast extract 1.00g
Sodium resazurin (0.1% w/v) 0.50ml
NaHCO3 4.50g
Na-acetate 1.40g
Na-butyrate 1.40g
Wolin's vitamin solution (10x) 1.00ml
Na2S x 9 H2O 0.40g
Distilled water 1000.00ml
Dissolve ingredients (except bicarbonate, acetate, butyrate, vitamins, and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add acetate, butyrate, vitamins, and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Before use, check the complete medium's pH and adjust it to 7.0 - 7.2, if necessary.
For DSM 2925: Replace acetate and butyrate with 1.00 g/l ethanol absolute added from a sterile anoxic stock solution after autoclaving.
For DSM 7213: Replace acetate and butyrate with 0.60 g/l Na-benzoate added after autoclaving from an anoxic stock solution sterilized by filtration.
For DSM 100382: Replace acetate and butyrate with 1.60 g/l Na-fumarate added after autoclaving from an anoxic stock solution sterilized by filtration.
Trace element solution SL-10 (from medium 320)
HCl (25%) 10.00ml，F(xiàn)eCl2 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 24.00mgNa2MoO4 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.
Selenite-tungstate solution (from medium 385)
NaOH 0.50g，Na2SeO3 x 5 H2O 3.00mg，Na2WO4 x 2 H2O 4.00mg，
Distilled water 1000.00ml
Wolin's vitamin solution (10x) (from medium 120)
Biotin 20.00mg，F(xiàn)olic acid 20.00mg，Pyridoxine hydrochloride 100.00mg，Thiamine HCl 50.00mg，
Riboflavin 50.00mg，Nicotinic acid 50.00mg，Calcium D-(+)-pantothenate 50.00mg，Vitamin B12 1.00mg，
p-Aminobenzoic acid 50.00mg，(DL)-alpha-Lipoic acid 50.00mg，Distilled water 1000.00ml
