Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 5.55 | g | ||
|
MgCl2 x 6 H2O | 2.29 | g | ||
|
CaCl2 x 2 H2O | 0.34 | g | ||
|
KCl | 0.15 | g | ||
|
Na2SO4 | 0.91 | g | ||
|
KNO3 | 1.01 | g | ||
|
HEPES buffer
(SIGMA) |
2.38 | g | ||
|
NH4Cl | 0.05 | g | ||
|
SrCl2
(0.1% w/v) |
8.00 | ml | ||
|
H3BO3
(0.1% w/v) |
6.00 | ml | ||
|
NaF
(0.1% w/v) |
4.00 | ml | ||
|
K2HPO4
(0.1% w/v) |
1.00 | ml | ||
|
Trace element solution SL-10 | 1.00 | ml | ||
|
Selenite-tungstate solution | 1.00 | ml | ||
|
NaHCO3 | 0.20 | g | ||
|
Na2S2O3 x 5 H2O | 2.48 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Distilled water | 980.00 | ml | ||
1 Dissolve ingredients (except bicarbonate, thiosulfate and vitamins) and adjust pH to 7.3. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes) and autoclave. Add thiosulfate and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of vitamins and thiosulftae should be sterilized by filtration. Adjust pH of the complete medium to 7.3 - 7.5, if necessary. | |||||
2 After inoculation pressurize vials to 0.7 bar overpressure with sterile 100% N2 gas. |
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 | |
---|---|---|---|
|
NaOH | 0.5 | g |
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
Distilled water | 1000.0 | 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 |