1478 .
NATRANAEROARCHAEUM MEDIUM AARC1
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Equipment needed:
Autoclave
Filter
Hungate tubes
Final pH:
3.0 - 4.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 72.000 | g | ||
|
|
K2HPO4 | 2.500 | g | ||
|
|
(NH4)2SO4 | 0.125 | g | ||
|
|
Na2CO3 | 142.500 | g | ||
|
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NaHCO3 | 22.500 | g | ||
|
|
Trace element solution | 1.000 | ml | ||
|
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Sulfur powder | 2.000 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Prepare the liquid medium. Dispense the medium into Hungate tubes or serum bottles under gassing with N2. Steam medium for 3h on each of 3 successive days. Before inoculation add from sterile stock solutions: | |||||
|
|
MgCl2 x 6 H2O | 1.200 | mM | ||
|
|
NaHCO3 | 1.250 | mM | ||
|
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NH4Cl | 3.000 | mM | ||
|
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Sodium formate | 50.000 | mM | ||
|
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Sodium acetate | 2.000 | mM | ||
|
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Yeast extract | 20.000 | mg/l | ||
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Vitamine mixture | 1.000 | ml | ||
|
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Disodium sulfide
(1M filter sterilized stock) |
1.000 | mM | ||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
|
FeSO4 x 7 H2O | 2.00 | g | ||
|
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ZnSO4 | 0.10 | g | ||
|
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MnCl2 | 0.03 | g | ||
|
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H3BO3 | 0.30 | g | ||
|
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CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 5 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.02 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 3.0-4.0 with HCl. | |||||
| 2 Autoclave at 121 °C for 15 min. | |||||
| 3 Note: autoclave in closed bottles | |||||
Vitamine mixture #
400
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Acidic thiamine / Ca pantothenate solution | 100 | ml | ||
|
|
Basic vitamin solution | 100 | ml | ||
|
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Neutral folic acid / riboflavin solution | 100 | ml | ||
|
|
Neutral B12 solution 0.005 g/L | 100 | ml | ||
| 1 Sterilize by filtration. | |||||
| 2 Mix the four solutions in the ratio 1:1:1:1. | |||||
Acidic thiamine / Ca pantothenate solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine | 5 | mg |
|
|
Calcium pantothenate | 5 | mg |
|
|
Distilled water
(acidified to pH 3.0 with HCl) |
100 | ml |
Basic vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2 | mg |
|
|
PABA
(p-aminobenzoic acid) |
5 | mg |
|
|
Nicotinic acid | 5 | mg |
|
|
Pyridoxine | 10 | mg |
|
|
NaOH
(0.1N) |
100 | ml |
Neutral folic acid / riboflavin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Folic acid | 2 | mg |
|
|
Riboflavin | 5 | mg |
|
|
Distilled water | 100 | ml |