DSMZ 500 . CLOSTRIDIUM (GV) MEDIUM

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Equipment needed: Autoclave Filter Gassing station Hungate tubes

Final gas composition: 80 % N2 20 % CO2 read more

Final pH: 7.0 - 7.2

Select a strain (optional):

ml
Compound Amount Unit
K2HPO4 0.35 g
KH2PO4 0.23 g
NH4Cl 0.50 g
NaCl 2.25 g
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
2.00 ml
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Yeast extract 1.00 g
Sodium resazurin
(0.1% w/v)
0.50 ml
NaHCO3 3.00 g
MgSO4 x 7 H2O 0.50 g
CaCl2 x 2 H2O 0.25 g
D-Glucose 5.00 g
Wolin's vitamin solution (10x) 1.00 ml
L-Cysteine HCl x H2O 0.30 g
Na2S x 9 H2O 0.30 g
Distilled water 1000.00 ml
1 Dissolve ingredients (except bicarbonate, magnesium sulfate, calcium chloride, glucose, vitamins, cysteine and sulfide), then 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, dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add magnesium sulfate, calcium chloride, glucose, cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2, if necessary.

Trace element solution SL-10 #

1000
Compound Amount Unit
HCl
(25%)
10.0 ml
FeCl2 x 4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2 x 4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2 x 6 H2O 190.0 mg
CuCl2 x 2 H2O 2.0 mg
NiCl2 x 6 H2O 24.0 mg
Na2MoO4 x 2 H2O 36.0 mg
Distilled water 990.0 ml
1 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 #

1000
Compound Amount Unit
NaOH 0.5 g
Na2SeO3 x 5 H2O 3.0 mg
Na2WO4 x 2 H2O 4.0 mg
Distilled water 1000.0 ml