785 .
NAD S1 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| 1 Mix components thoroughly, distribute the medium into culture vessels (e.g., 5 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add per liter the following solutions from anaerobic stocks (autoclaved or *filter-sterilized): | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.3 | g | ||
|
|
KH2PO4 | 0.3 | g | ||
|
|
KCl | 0.5 | g | ||
|
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
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NaCl | 15.0 | g | ||
|
|
KNO3 | 1.5 | g | ||
|
|
Yeast extract | 0.5 | g | ||
|
|
Trace minerals | 10.0 | ml | ||
|
|
Distilled water | 1000.0 | ml | ||
|
|
NaHCO3
(*, 8%) |
25.0 | ml | ||
|
|
MgCl2 x 6 H2O
(10%) |
5.0 | ml | ||
|
|
Glucose
(1 M) |
20.0 | ml | ||
|
|
Trace vitamins | 10.0 | ml | ||
Trace minerals #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
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MnSO4 x n H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 | 0.01 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. | |||||
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 |