454 .
EBA MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
7.2 - 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.20 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
MgSO4 x 7 H2O | 0.50 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Na2SeO3 x 5 H2O
(0.26 g/ 1000 ml 10 mM NaOH) |
1.00 | ml | ||
|
|
Glycine | 2.00 | g | ||
|
|
Distilled water | 950.00 | ml | ||
| 1 pH 7.2 to 7.4 | |||||
| 2 Before inoculation add per 100 ml of medium: | |||||
|
|
Biotin
(10 mg/l, autoclave for 10 minutes under N2) |
0.20 | ml | ||
|
|
NaHCO3
(84 g/l, autoclaved under CO2) |
3.00 | ml | ||
|
|
Na2S
(60 g/l, autoclaved under N2) |
1.00 | ml | ||
| 3 Prepare and distribute medium anaerobically by using the Hungate technique. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
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 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||