348 .
METHANOHALOPHILUS PORTUCALENSIS MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
7.3 - 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 120.00 | g | ||
|
|
MgCl2 x 6 H2O | 7.00 | g | ||
|
|
MgSO4 x 7 H2O | 6.00 | g | ||
|
|
CaCl2 x 2 H2O | 0.50 | g | ||
|
|
K2HPO4 x 3 H2O | 0.40 | g | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
KCl | 3.80 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
Yeast extract
(OXOID) |
2.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
2.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Na2CO3 | 1.50 | g | ||
|
|
Trimethylamine-HCl | 1.90 | g | ||
|
|
Na2S x 9 H2O | 0.25 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except cysteine, carbonate, trimethylamine and sulfide, bring medium to the boil, then cool to room temperature while sparging with 80% N2 and 20% CO2 gas mixture. Add cysteine and adjust pH to 6.5, then dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add trimethylamine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.3 - 7.4, 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. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |