671 .
THERMOANAEROBIUM MEDIUM
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Equipment needed:
Autoclave
Final pH:
7.3
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract | 3.00 | g | ||
|
|
Tryptone
(BD-Difco) |
10.00 | g | ||
|
|
NH4Cl | 0.90 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
KH2PO4 | 0.75 | g | ||
|
|
K2HPO4 | 1.50 | g | ||
|
|
Trace element solution | 9.00 | ml | ||
|
|
Trace vitamins | 5.00 | ml | ||
|
|
FeSO4 x 7 H2O | 3.00 | mg | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 950.00 | ml | ||
| 1 Mix ingredients and adjust pH to 7.0. Distribute the medium into culture vessels under a N2 atmosphere, seal with butyl rubber stoppers and autoclave. Separately autoclave the following solutions under a N2 atmosphere and add to the medium (per 950 ml): | |||||
|
|
Glucose
(10%) |
50.00 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
10.00 | ml | ||
| 2 Readjust pH of the medium to 7.2 - 7.4 with sterile NaOH, if necessary. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 12.800 | g | ||
|
|
FeCl2 x 4 H2O | 0.200 | g | ||
|
|
MnCl2 x 4 H2O | 0.100 | g | ||
|
|
CoCl2 x 6 H2O | 0.170 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
ZnCl2 | 0.100 | g | ||
|
|
CuCl2 x 2 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.010 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.010 | g | ||
|
|
NiCl2 x 6 H2O | 0.026 | g | ||
|
|
NaCl | 1.000 | g | ||
|
|
Na2SeO3 | 0.020 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Dissolve nitrilotriacetic acid first in 200 ml of distilled water, and adjust pH 6.5 with KOH. Then add the remaining salts and adjust volume to 1000 ml. | |||||
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |