1090 .
MMJS MEDIUM 2
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Equipment needed:
Autoclave
Filter
Final pH:
6.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 20.00 | g | ||
|
|
K2HPO4 | 0.14 | g | ||
|
|
CaCl2 x 2 H2O | 1.00 | g | ||
|
|
MgSO4 x 7 H2O | 3.40 | g | ||
|
|
MgCl2 x 6 H2O | 4.18 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
NiCl2 x 6 H2O | 0.50 | mg | ||
|
|
Na2SeO3 x 5 H2O | 0.50 | mg | ||
|
|
Na2WO4 x 2 H2O | 0.10 | mg | ||
|
|
(NH4)2Ni(SO4)2 x 6 H2O | 10.00 | mg | ||
|
|
Trace minerals | 10.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components and adjust pH to 5.5. Distribute the medium into culture vessels (e.g., 20 ml in 60 ml-serum bottles) under a stream of N2, seal with butyl rubber stoppers and add Autoclave. After cooling, add per liter the following solutions (filter-sterilized): | |||||
|
|
Trace vitamins | 1.00 | ml | ||
|
|
NaHCO3
(8%) |
3.75 | ml | ||
|
|
Na2S2O3
(1 M) |
5.00 | ml | ||
| 2 Replace the gas phase with N2-CO2-O2 (94:5:1, v/v/v, 100 kPa). Check final pH to be around 6.2. | |||||
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. | |||||
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 |