1515 .
OCEANIRHABDUS MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
6.5 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2SO4 | 1.0 | g | ||
|
|
NaCl | 30.0 | g | ||
|
|
KCl | 0.7 | g | ||
|
|
MgCl2 x 6 H2O | 5.0 | g | ||
|
|
CaCl2 x 2 H2O | 1.0 | g | ||
|
|
K2HPO4 | 0.4 | g | ||
|
|
KH2PO4 | 0.4 | g | ||
|
|
NH4Cl | 0.5 | g | ||
|
|
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v) |
2.0 | ml | ||
|
|
Trace element solution SL-10 | 1.0 | ml | ||
|
|
Yeast extract
(OXOID) |
3.0 | g | ||
|
|
Tryptone
(BD Bacto) |
4.0 | g | ||
|
|
Na-acetate | 6.8 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.6 | g | ||
|
|
NaHCO3 | 2.5 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except cysteine and bicarbonate, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve cysteine, adjust pH to 6.5, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 6.5 - 7.0, if necessary. | |||||
Trace element solution SL-10 #
1000
| 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. | |||||