1307 .
PPLL MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.00 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
Na2SO4 | 4.00 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KH2PO4 | 0.20 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
Yeast extract | 0.50 | g | ||
|
|
Sodium malate | 0.89 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components and autoclave. Cool down under a N2 gas stream. Aseptically and anaerobically add per liter the following solutions from sterile anaerobic stocks : | |||||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
NaHCO3
(8%) |
30.00 | ml | ||
|
|
Vitamin solution | 1.00 | ml | ||
|
|
Thiamine solution | 1.00 | ml | ||
|
|
Vitamin B12 solution 0.05 g/L | 1.00 | ml | ||
| 2 Adjust pH to 7.0-7.3. Aseptically and anaerobically distribute the medium into culture vessels (e.g., 10 ml medium in Balch tubes), fill the head space with a gas mixture of N2-CO2 (4:1, v/v), and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution. | |||||
|
|
Na2S x 9 H2O
(5%) |
5.00 | ml | ||
|
|
L-Cysteine HCl x H2O
(5%) |
5.00 | ml | ||
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(25%) |
12.5 | ml |
|
|
FeSO4 x 7 H2O | 2.1 | g |
|
|
H3BO3 | 30.0 | mg |
|
|
MnCl2 x 4 H2O | 100.0 | mg |
|
|
CoCl2 x 6 H2O | 190.0 | mg |
|
|
NiCl2 x 6 H2O | 24.0 | mg |
|
|
CuCl2 x 2 H2O | 2.0 | mg |
|
|
ZnSO4 x 7 H2O | 144.0 | mg |
|
|
Na2MoO4 x 2 H2O | 36.0 | mg |
|
|
Distilled water | 987.0 | ml |
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
p-Aminobenzoic acid | 4 | mg |
|
|
Biotin | 1 | mg |
|
|
Nicotinic acid | 10 | mg |
|
|
DL-Calcium pantothenate | 5 | mg |
|
|
Pyridoxine hydrochloride | 10 | mg |
|
|
Sodium phosphate buffer
(10 mM, pH 7.1) |
100 | ml |
Thiamine solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine HCl | 10 | mg |
|
|
Sodium phosphate buffer
(25 mM, pH 3.4) |
100 | ml |