1312 .
MODIFIED MV MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| 1 Mix ingredients, bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream and autoclave under the same gas mixture. After cooling, aseptically and anaerobically add per liter the following solutions (filter-sterilized): | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.000 | g | ||
|
|
NaCl | 2.100 | g | ||
|
|
MgCl2 x 6 H2O | 0.825 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
KH2PO4 | 0.500 | g | ||
|
|
Trace element solution SL-10 | 1.000 | ml | ||
|
|
Selenite-tungstate solution | 1.000 | ml | ||
|
|
Yeast extract
(BD-Difco) |
1.000 | g | ||
|
|
Resazurin
(0.1%) |
0.500 | ml | ||
|
|
Trace vitamins | 10.000 | ml | ||
|
|
NaHCO3
(8%) |
30.000 | ml | ||
|
|
Taurine
(0.5 M) |
20.000 | ml | ||
|
|
Sodium pyruvate
(1 M) |
20.000 | ml | ||
|
|
Sodium L-lactate
(0.5 M) |
20.000 | ml | ||
|
|
Hemin solution | 10.000 | ml | ||
| 2 Prior to inoculation, aseptically and anaerobically add 0.01 volume of L-cysteine x HCl x H2O to the medium (3% solution, autoclaved and stored under a N2 atmosphere). | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%, 7.7 M) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | 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 |