695 .
NAUTILIA ABYSSI MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 25.00 | g | ||
|
|
NaNO3 | 1.00 | g | ||
|
|
NH4Cl | 0.33 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
CaCl2 x 2 H2O | 0.33 | g | ||
|
|
MgCl2 x 6 H2O | 0.33 | g | ||
|
|
MES | 1.95 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Trace minerals | 10.00 | ml | ||
|
|
Sulfur
(powder) |
12.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components except sulfur, adjust pH to 6.0 and autoclave under a N2 atmosphere. Distribute the medium into culture vessels (e.g., 20 ml in 100 ml serum bottles) containing appropriate amount of sulfur under a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized) into the medium (per liter): | |||||
|
|
Trace vitamins (see Medium No. 197)
(*) |
10.00 | ml | ||
|
|
KH2PO4
(2%) |
20.00 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
10.00 | ml | ||
| 2 Pressurize the inoculated culture vessel to 200 kPa H2-CO2 (4:1, v/v). | |||||
Trace minerals #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
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. | |||||