1093 .
MJ BASAL MEDIUM WITH THIOSULFATE/2
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Equipment needed:
Autoclave
Filter
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MJ(-N) synthetic seawater | 1000.00 | ml | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KNO3 | 0.25 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components thoroughly and autoclave. After cooling, add the following solutions (filter-sterilized): | |||||
|
|
NaHCO3
(8%) |
10.00 | ml | ||
|
|
Na2S2O3 x 5 H2O
(10%) |
15.00 | ml | ||
|
|
Trace vitamins | 1.00 | ml | ||
| 2 Aseptically distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) and seal with butyl rubber stoppers. Replace the gas phase with a N2-CO2 (80:20, v/v) gas mixture, and pressurize to 200 kPa. Adjust pH to 6.0, if necessary. | |||||
MJ(-N) synthetic seawater #
1010
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 30.00 | g |
|
|
K2HPO4 | 0.14 | g |
|
|
CaCl2 x 2 H2O | 0.14 | g |
|
|
MgSO4 x 7 H2O | 3.40 | g |
|
|
MgCl2 x 6 H2O | 4.18 | g |
|
|
KCl | 0.33 | g |
|
|
NiCl2 x 6 H2O | 0.50 | mg |
|
|
Na2SeO3 x 5 H2O | 0.50 | mg |
|
|
Fe(NH4)2(SO4)2 x 6 H2O | 0.01 | g |
|
|
Modified Wolin's mineral solution | 10.00 | ml |
|
|
Distilled water | 1000.00 | ml |
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | 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. | |||||