| |
Bacto Tryptone (Difco) |
5 |
g |
| |
Bacto Peptone (Difco) |
5 |
g |
| |
Bacto Yeast Extract (Difco) |
10 |
g |
| |
Bacto Beef Extract (Difco) |
5 |
g |
| |
Glucose |
5 |
g |
| |
K2HPO4 |
2 |
g |
| |
Cysteine-HCl·H2O |
0.5 |
g |
| |
Tween 80 |
1 |
ml |
| |
Resazurin |
1 |
mg |
| |
Salt solution* |
40 |
ml |
| |
Hemin solution** |
10 |
ml |
| |
Menadione solution*** |
10 |
ml |
| |
Distilled water |
940 |
ml |
| |
pH 7.2 |
|
| |
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|
Dissolve ingredients except cysteine-HCl, hemin and menadione solutions, and adjust the pH to 7.2. Dispense the medium into suitable culture vessels under a stream of N2 gas and seal with butyl rubber stoppers. After autoclaving (121°C, 15 min), aseptically and anaerobically add the other ingredients. |
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*Salt solution |
| |
|
|
CaCl2·2H2O |
0.25 |
g |
| |
|
|
MgSO4·7H2O |
0.5 |
g |
| |
|
|
K2HPO4 |
1 |
g |
| |
|
|
KH2PO4 |
1 |
g |
| |
|
|
NaHCO3 |
10 |
g |
| |
|
|
NaCl |
2 |
g |
| |
|
|
Distilled water |
1 |
L |
| |
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|
**Hemin solution |
| |
|
Dissolve 50 mg hemin first in 1 ml of 1 N NaOH. Then adjust volume to 100 ml. |
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***Menadione solution |
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|
Dissolve 5 mg menadione first in 1 ml of ethanol. Then adjust volume to 100 ml. |