Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 0.33 | g | ||
|
KCl | 0.33 | g | ||
|
KH2PO4 | 0.33 | g | ||
|
MgCl2 x 6 H2O | 0.33 | g | ||
|
CaCl2 x 2 H2O | 0.33 | g | ||
|
Na2SO4 | 2.80 | g | ||
|
Trace element solution SL-10 | 1.00 | ml | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Yeast extract | 3.00 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Neutralized sulfide solution 3% (w/v) | 15.00 | ml | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except yeast extract, vitamins and sulfide. Adjust pH to 4.5 with H2SO4 and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add yeast extract, vitamins (sterilized by filtration), and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The sulfide stock solution (3% w/v) should be neutralized before use (see medium 28). The pH of the complete medium should be at 4.5. |
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. |
Compound | Amount | Unit | |
---|---|---|---|
|
Biotin | 20 | mg |
|
Folic acid | 20 | mg |
|
Pyridoxine hydrochloride | 100 | mg |
|
Thiamine HCl | 50 | mg |
|
Riboflavin | 50 | mg |
|
Nicotinic acid | 50 | mg |
|
Calcium D-(+)-pantothenate | 50 | mg |
|
Vitamin B12 | 1 | mg |
|
p-Aminobenzoic acid | 50 | mg |
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
Distilled water | 1000 | ml |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Na2S x 9 H2O | 3 | g | ||
|
Distilled water | 100 | ml | ||
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle. |