| |
Hipolypepton* |
10 |
g |
| |
Bacto Yeast Extract (Difco) |
2.5 |
g |
| |
Glucose |
2 |
g |
| |
Maltose |
2 |
g |
| |
Cellobiose |
2 |
g |
| |
MgSO4·7H2O |
0.045 |
g |
| |
CaCl2·2H2O |
0.09 |
g |
| |
K2HPO4 |
0.45 |
g |
| |
KH2PO4 |
0.45 |
g |
| |
NaCl |
0.9 |
g |
| |
NaHCO3 |
4 |
g |
| |
Vitamin solution** |
10 |
ml |
| |
Hemin solution*** |
10 |
ml |
| |
Resazurin |
1 |
mg |
| |
Cysteine-HCl |
1 |
g |
| |
Distilled water |
1 |
L |
| |
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*FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan. |
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Mix ingredients except for vitamin and hemin solutions, and cysteine-HCl. Dispense the medium into suitable culture vessels and autoclave under a N2 atmosphere. Separately autoclave hemin solution and cystein-HCl (5% solution) under a N2 atmosphere. Sterilize vitamin solution by filatration. Prior to inocluation, aseptically and anaerobically add vitamin, hemin and cysteine-HCl solutions. |
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**Vitamin solution |
| |
|
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Biotin |
2 |
mg |
| |
|
|
Folic acid |
2 |
mg |
| |
|
|
Pyridoxine-HCl |
10 |
mg |
| |
|
|
Thiamine-HCl |
5 |
mg |
| |
|
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Riboflavin |
5 |
mg |
| |
|
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Nicotinic acid |
5 |
mg |
| |
|
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Ca-pantothenate |
5 |
mg |
| |
|
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p-Aminobenzoic acid |
1 |
mg |
| |
|
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Vitamin B12 |
0.01 |
mg |
| |
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Distilled water |
1 |
L |
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***Hemin solution |
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Dissolve 50 mg hemin first in 1 ml of 1 N NaOH. Then adjust volume to 100 ml. |
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