1615 .
Basic mineral medium
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Equipment needed:
Autoclave
Filter
Final pH:
7.0 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
20 mM K-P-Buffer | 1000.0 | ml | ||
| 1 After sterilization add from sterile stock solutions: | |||||
|
|
MgSO4 x 7 H2O
(1 M) |
1.0 | ml | ||
|
|
Sodium acetate
(1 M) |
10.0 | ml | ||
|
|
CaCl2 x 2 H2O
(5%) |
1.0 | ml | ||
| 2 might result in colloid formation, especially when added into hot medium, but dissolves after vigorous shaking | |||||
|
|
Na-aspartate
(2 M) |
1.0 | ml | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
L-Methionine
(50 mM, filter sterilized, neutralised) |
1.0 | ml | ||
|
|
Vitamine mixture | 0.5 | ml | ||
| 3 pH 7.0-7.2 | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
|
FeSO4 x 7 H2O | 2.00 | g | ||
|
|
ZnSO4 | 0.10 | g | ||
|
|
MnCl2 | 0.03 | g | ||
|
|
H3BO3 | 0.30 | g | ||
|
|
CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 5 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.02 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 3.0-4.0 with HCl. | |||||
| 2 Autoclave at 121 °C for 15 min. | |||||
| 3 Note: autoclave in closed bottles | |||||
20 mM K-P-Buffer #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K2HPO4
(0.1 M) |
130 | ml | ||
|
|
KH2PO4
(0.1 M) |
70 | ml | ||
|
|
Distilled water | 800 | ml | ||
| 1 Sterilize at 121°C for 20 minutes in 1L Schott bottle | |||||
Vitamine mixture #
400
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Acidic thiamine / Ca pantothenate solution | 100 | ml | ||
|
|
Basic vitamin solution | 100 | ml | ||
|
|
Neutral folic acid / riboflavin solution | 100 | ml | ||
|
|
Neutral B12 solution 0.005 g/L | 100 | ml | ||
| 1 Sterilize by filtration. | |||||
| 2 Mix the four solutions in the ratio 1:1:1:1. | |||||
Acidic thiamine / Ca pantothenate solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine | 5 | mg |
|
|
Calcium pantothenate | 5 | mg |
|
|
Distilled water
(acidified to pH 3.0 with HCl) |
100 | ml |
Basic vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2 | mg |
|
|
PABA
(p-aminobenzoic acid) |
5 | mg |
|
|
Nicotinic acid | 5 | mg |
|
|
Pyridoxine | 10 | mg |
|
|
NaOH
(0.1N) |
100 | ml |
Neutral folic acid / riboflavin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Folic acid | 2 | mg |
|
|
Riboflavin | 5 | mg |
|
|
Distilled water | 100 | ml |