|
NaCl |
30 |
g |
|
KCl |
0.33 |
g |
|
NH4Cl |
0.25 |
g |
|
MgCl2·6H2O |
2.8 |
g |
|
MgSO4·7H2O |
3.4 |
g |
|
FeSO4·7H2O |
0.025 |
g |
|
K2HPO4 |
0.3 |
g |
|
NaBr |
0.01 |
g |
|
Trace minerals* |
10 |
ml |
|
Vitamin solution** |
10 |
ml |
|
Bacto Yeast Extract (Difco) |
3 |
g |
|
Bacto Tryptone (Difco) |
3 |
g |
|
PIPES |
6 |
g |
|
Sulfur (powder) |
10 |
g |
|
Resazurin |
1 |
mg |
|
Na2S·9H2O |
0.5 |
g |
|
Distilled water |
1 |
L |
|
|
|
Mix ingredients except vitamin solution, sulfur and Na2S·9H2O, and adjust pH to 7. Autoclave under a N2 atmosphere. Filter-sterilize vitamin solution. Sulfur is sterilized separately by autoclaving for 60 min at 105℃ on each of 3 successive days. Neutralize Na2S·9H2O as 5% solution and autoclave under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add vitamin, sulfur and Na2S·9H2O. |
|
|
|
|
|
|
|
|
|
*Trace minerals |
|
|
|
Nitrilotriacetic acid |
1.5 |
g |
|
|
|
MgSO4·7H2O |
3 |
g |
|
|
|
MnSO4·xH2O |
0.5 |
g |
|
|
|
NaCl |
1 |
g |
|
|
|
FeSO4·7H2O |
0.1 |
g |
|
|
|
CoSO4·7H2O |
0.1 |
g |
|
|
|
CaCl2·2H2O |
0.1 |
g |
|
|
|
ZnSO4·7H2O |
0.1 |
g |
|
|
|
CuSO4·5H2O |
0.01 |
g |
|
|
|
AlK(SO4)2 |
0.01 |
g |
|
|
|
H3BO3 |
0.01 |
g |
|
|
|
Na2MoO4·2H2O |
0.01 |
g |
|
|
|
Distilled water |
1 |
L |
|
|
|
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. |
|
|
|
|
|
|
|
|
|
** Vitamin solution |
|
|
|
Biotin |
2 |
mg |
|
|
|
Folic acid |
2 |
mg |
|
|
|
Pyridoxine-HCl |
10 |
mg |
|
|
|
Thiamine-HCl |
5 |
mg |
|
|
|
Riboflavin |
5 |
mg |
|
|
|
Nicotinic acid |
5 |
mg |
|
|
|
Ca-pantothenate |
5 |
mg |
|
|
|
Vitamin B12 |
0.1 |
mg |
|
|
|
p-Aminobenzoic acid |
5 |
mg |
|
|
|
Lipoic acid |
5 |
mg |
|
|
|
Distilled water |
1 |
L |
|