|
KH2PO4 |
0.2 |
g |
|
NH4Cl |
0.25 |
g |
|
NaCl |
1 |
g |
|
MgCl2·6H2O |
0.4 |
g |
|
KCl |
0.5 |
g |
|
CaCl2·2H2O |
0.15 |
g |
|
Sodium acetate |
0.16 |
g |
|
Trace element solution* |
2 |
ml |
|
Vitamin solution** |
2 |
ml |
|
Resazurin |
1 |
mg |
|
NaHCO3 |
4.8 |
g |
|
Cysteine-HCl |
0.5 |
g |
|
Na2S·9H2O |
0.5 |
g |
|
Distilled water |
1 |
L |
|
|
|
Mix ingredients except vitamin solution, NaHCO3, Na2S·9H2O and cysteine-HCl, and autoclave under a H2/CO2 (80/20) atmosphere. Separately autoclave cysteine-HCl and Na2S·9H2O as 5% solutions under a N2 atmosphere. Sterilize vitamin and 8% NaHCO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add vitamin, NaHCO3, cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated bottles to 150 kPa with H2/CO2. |
|
|
|
|
|
|
|
|
|
*Trace element solution |
|
|
|
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 |
|
|
|
H3BO3 |
0.01 |
g |
|
|
|
Na2MoO4·2H2O |
0.01 |
g |
|
|
|
NiCl2·6H2O |
0.025 |
g |
|
|
|
Na2SeO3·5H2O |
0.3 |
g |
|
|
|
Na2WO4·2H2O |
0.01 |
g |
|
|
|
Distiled water |
1 |
L |
|
|
|
Dissolve nitrilotoriacetic acid and adjust pH to 6.5 with KOH. Then 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 |
|
|
|
p-Aminobenzoic acid |
1 |
mg |
|
|
|
Vitamin B12 |
0.01 |
mg |
|
|
|
Distilled water |
1 |
L |
|