950 .
NH-JN4 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2SO4 | 1.0 | g | ||
|
|
NaCl | 30.0 | g | ||
|
|
KCl | 0.7 | g | ||
|
|
MgCl2 x 6 H2O | 5.0 | g | ||
|
|
CaCl2 x 2 H2O | 1.0 | g | ||
|
|
K2HPO4 | 0.4 | g | ||
|
|
KH2PO4 | 0.4 | g | ||
|
|
NH4Cl | 0.5 | g | ||
|
|
Fe(NH4)2(SO4)2 x 6 H2O | 2.0 | mg | ||
|
|
Yeast extract
(BD-Difco) |
3.0 | g | ||
|
|
Tryptone
(BD-Difco) |
4.0 | g | ||
|
|
Sodium acetate | 6.8 | g | ||
|
|
L-Cysteine HCl x H2O | 0.6 | g | ||
|
|
Trace element solution SL-10 | 1.0 | ml | ||
|
|
Resazurin | 1.0 | mg | ||
|
|
Distilled water | 960.0 | ml | ||
| 1 Mix components thoroughly and adjust pH to 6.8. Distribute the medium into culture vessels under a N2-CO2 (9:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add the following solution (per 960 ml) from anaerobic stock (filter-sterilized) | |||||
|
|
NaHCO3
(5%) |
40.0 | ml | ||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%, 7.7 M) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. | |||||