26 .
ACID RHODOSPIRILLACEAE MEDIUM
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Selected strain: Rhodomicrobium vannielii DSM 162
This strain uses the standard medium without modifications.
Growth conditions: 7-14 days
28 °C
Equipment needed:
Autoclave
Tungsten lamp
Final gas composition:
100 % N2
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Final pH:
5.7
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract | 0.20 | g | ||
|
|
Disodium succinate | 1.00 | g | ||
|
|
Ammonium acetate | 0.50 | g | ||
|
|
Fe(III) citrate
(0.1% in H2O) |
5.00 | ml | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
MgSO4 x 7 H2O | 0.40 | g | ||
|
|
NaCl | 0.40 | g | ||
|
|
NH4Cl | 0.40 | g | ||
|
|
CaCl2 x 2 H2O | 0.05 | g | ||
|
|
Vitamin B12
(10 mg in 100 ml H2O) |
0.40 | ml | ||
|
|
Trace element solution SL-6 | 1.00 | ml | ||
|
|
L-Cysteine HCl | 0.30 | g | ||
|
|
Resazurin
(0,1%) |
0.50 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 5.7. | |||||
| 2 Boil the medium for a few minute. | |||||
| 3 Sparge medium with 100% N2 gas mixture to make it anoxic. | |||||
| 4 Fill 10 ml in 15 ml tubes with a rubber septum under a stream of nitrogen gas. | |||||
| 5 Autoclave at 121 °C for 20 min. | |||||
| 6 Sterile syringes are used to inoculate and remove samples. | |||||
| 7 Incubate in the light using a tungsten lamp. | |||||
Trace element solution SL-6 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnSO4 x 7 H2O | 0.10 | g |
|
|
MnCl2 x 4 H2O | 0.03 | g |
|
|
H3BO3 | 0.30 | g |
|
|
CoCl2 x 6 H2O | 0.20 | g |
|
|
CuCl2 x 2 H2O | 0.01 | g |
|
|
NiCl2 x 6 H2O | 0.02 | g |
|
|
Na2MoO4 x 2 H2O | 0.03 | g |
|
|
Distilled water | 1000.00 | ml |