244 .
JCM MEDIUM No. 244
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Selected strain: Thermococcus aegaeus DSM 12767
This strain uses the standard medium without modifications.
Equipment needed:
Autoclave
Filter
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Synthetic seawater (2 x)
(2 x) |
500.00 | ml | ||
|
|
NaHCO3 | 0.16 | g | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
Trace minerals | 15.00 | ml | ||
|
|
Ni-Se-W solution | 10.00 | ml | ||
|
|
Bacto peptone
(BD-Difco) |
2.00 | g | ||
|
|
Yeast extract
(BD-Difco) |
1.00 | g | ||
|
|
Resazurin | 0.75 | mg | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Sulfur
(powder) |
10.00 | g | ||
|
|
Distilled water | 500.00 | ml | ||
| 1 Mix ingredients, except sulfur and Na2S x 9H2O, and adjust pH to 6.0 with H2SO4. Sterilize the medium by filtration. Steam sulfur for 3 hr on each of 3 successive days. Neutralize Na2S x 9H2O as a 5% solution, and autoclave under a N2 atmosphere. Distribute the medium and sulfur into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S x 9H2O solution, and readjust pH to 6.0 with sterile 1 N H2SO4. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v). | |||||
Synthetic seawater (2 x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 55.40 | g |
|
|
MgSO4 x 7 H2O | 14.00 | g |
|
|
MgCl2 x 6 H2O | 11.00 | g |
|
|
KCl | 1.30 | g |
|
|
NaBr | 0.20 | g |
|
|
H3BO3 | 0.06 | g |
|
|
SrCl2 x 6 H2O | 0.03 | g |
|
|
KI | 0.10 | mg |
|
|
Trisodium citrate | 20.00 | mg |
|
|
CaCl2 x 2 H2O | 1.50 | g |
|
|
Distilled water | 1000.00 | ml |
Ni-Se-W solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NiCl2 x 6 H2O | 25.0 | mg |
|
|
(NH4)2Ni(SO4)2 x 6 H2O | 2.0 | g |
|
|
Na2SeO3 | 0.3 | mg |
|
|
Na2SeO4 | 10.0 | mg |
|
|
Na2WO4 x 2 H2O | 10.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Trace minerals #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x n H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 | 0.01 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. | |||||