151 .
PYROCOCCUS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Salt base solution | 1000.0 | ml | ||
|
|
Trace minerals | 10.0 | ml | ||
|
|
Citric acid | 5.0 | mg | ||
|
|
Yeast extract
(BD-Difco) |
1.0 | g | ||
|
|
Bacto peptone
(BD-Difco) |
5.0 | g | ||
|
|
Sulfur
(powder) |
30.0 | g | ||
|
|
Na2S x 9 H2O | 0.5 | g | ||
|
|
Resazurin | 1.0 | mg | ||
| 1 Mix ingredients, except sulfur and Na2S x 9H2O, and adjust pH to 6.5 with H2SO4. Sterilize by filtration under a N2 atmosphere. 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. Aseptically and anaerobically add sterile sulfur to the medium. Prior to inoculation, add the sterile Na2S x 9H2O solution, and readjust pH to 6.5, if necessary. | |||||
Salt base solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 13.850 | g |
|
|
MgSO4 x 7 H2O | 3.500 | g |
|
|
MgCl2 x 6 H2O | 2.750 | g |
|
|
KCl | 0.325 | g |
|
|
NaBr | 0.050 | g |
|
|
H3BO3 | 15.000 | mg |
|
|
SrCl2 x 6 H2O | 7.500 | mg |
|
|
KI | 0.050 | mg |
|
|
CaCl2 | 0.750 | g |
|
|
KH2PO4 | 0.500 | g |
|
|
(NH4)2Ni(SO4)2 x 6 H2O | 2.000 | mg |
|
|
Distilled water | 1000.000 | 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. | |||||