1131 .
HYDROGENIVIRGA OKINAWENSIS MEDIUM
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* Medium for Thiolapillus brandeum DSM 23672
apply 2 bar H2/CO2 (80:20)
Growth conditions: 2-3 days
30 °C
Equipment needed:
Autoclave
Filter
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Modified MJ synthetic seawater | 1000 | ml | ||
| 1 Sparge medium with 100% N2 gas mixture for 10 min to make anoxic. | |||||
| 2 Adjust pH to 7.0 with NaOH. | |||||
| 3 Sparge medium with 80% N2 and 20% CO2 gas mixture for 10 min. | |||||
|
|
Sulfur
(powder) |
3 | g/l | ||
| 4 Dispense the medium (10 mL) into 100 mL serum bottles, containing 0,03g sulfur (powder) under a stream of H2/CO2 (80:20) and seal with butyl rubber stoppers. | |||||
| 5 Autoclave at 121 °C for 20 min. | |||||
| 6 Prior to inoculation, add vitamins, NaNO3, and NaHCO3 (from filter sterilized stock solutions under a N2 gas atmosphere) to the medium. | |||||
|
|
Vitamin solution | 10 | ml/l | ||
|
|
NaNO3 | 1 | g/l | ||
|
|
NaHCO3 | 1 | g/l | ||
Modified MJ synthetic seawater #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 30.00 | g |
|
|
K2HPO4 | 0.14 | g |
|
|
CaCl2 x 2 H2O | 0.50 | g |
|
|
NH4Cl | 0.50 | g |
|
|
MgSO4 x 7 H2O | 3.40 | g |
|
|
MgCl2 x 6 H2O | 4.18 | g |
|
|
KCl | 0.33 | g |
|
|
FeSO4 x 7 H2O | 0.01 | g |
|
|
NiCl2 x 6 H2O | 1.00 | mg |
|
|
Na2SeO3 x 5 H2O | 1.00 | mg |
|
|
Modified Wolin's mineral solution II | 10.00 | ml |
|
|
Distilled water | 1000.00 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine-HCl x 2 H2O | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
D-Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.500 | g | ||
|
|
MgSO4 x 7 H2O | 3.000 | g | ||
|
|
MnSO4 x H2O | 0.500 | g | ||
|
|
NaCl | 1.000 | g | ||
|
|
FeSO4 x 7 H2O | 0.100 | g | ||
|
|
CoSO4 x 7 H2O | 0.180 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
ZnSO4 x 7 H2O | 0.180 | g | ||
|
|
CuSO4 x 5 H2O | 0.010 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.010 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.010 | g | ||
|
|
NiCl2 x 6 H2O | 0.025 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.300 | mg | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||