1663 .
BSM medium
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
8.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 3.62 | g | ||
|
|
K2HPO4 | 0.26 | g | ||
|
|
(NH4)Cl | 0.08 | g | ||
|
|
NaNO3 | 0.85 | g | ||
|
|
Na2SO4 | 0.05 | g | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Wolin's vitamin solution | 10.00 | ml | ||
|
|
Na-DL-lactate | 1.12 | g | ||
|
|
Yeast extract | 0.20 | g | ||
|
|
Distilled water | 936.50 | ml | ||
|
|
NaHCO3
(8%) |
52.50 | ml | ||
| 1 Dissolve ingredients (except vitamins and trace elements) in distilled water; boil for 1- 2 minutes and cool down to room temperature under 100% N2 gas, dispense in culture vessels and autoclave. After sterilization add NaHCO3, vitamins, and trace elements from sterile anoxic stock solutions prepared under 100% N2. Adjust pH to 8.2 and add sodium sulfide (Na2S) from a sterile anoxic solution to achieve a final concentration of 100 µM prior inoculation. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%; 7.7 M) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
MgCl2 x 6 H2O | 3.0 | g | ||
|
|
CaCl2 x 2 H2O | 0.1 | 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 1000.0 ml. | |||||
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |