1151 .
DESULFOBACTERIUM NAPHTHALENIVOLANS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.00 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
KH2PO4 | 0.20 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
|
Na2SO4 | 1.42 | g | ||
|
|
FeCl2 solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
Resazurin | 0.40 | mg | ||
|
|
Distilled water | 970.00 | ml | ||
| 1 Mix components and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized): | |||||
|
|
Vitamin solution | 1.00 | ml | ||
|
|
NaHCO3
(8%) |
30.00 | ml | ||
| 2 Aseptically and anaerobically distribute the medium into culture vessels (e.g., 50 ml of the medium in 120 ml serum bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, add per liter the following solution (autoclaved and stored under a N2 atmosphere): | |||||
|
|
Naphthalene solution
(1.5%) |
20.00 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
4.00 | ml | ||
Naphthalene solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Naphthalene | 1.5 | g |
|
|
2,2,4,4,6,8,8-Heptamethylnonane | 100.0 | ml |
FeCl2 solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
|
FeCl2 x 4 H2O | 1.5 | g |
|
|
Distilled water | 990.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnCl2 | 70 | mg |
|
|
MnCl2 x 4 H2O | 100 | mg |
|
|
H3BO3 | 6 | mg |
|
|
CoCl2 x 6 H2O | 190 | mg |
|
|
CuCl2 x 2 H2O | 2 | mg |
|
|
NiCl2 x 6 H2O | 24 | mg |
|
|
Na2MoO4 x 2 H2O | 36 | mg |
|
|
Distilled water | 1000 | ml |
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 20 | mg | ||
|
|
Nicotinic acid | 200 | mg | ||
|
|
p-Aminobenzoic acid | 100 | mg | ||
|
|
Thiamine HCl | 200 | mg | ||
|
|
DL-Calcium pantothenate | 100 | mg | ||
|
|
Pyridoxine hydrochloride | 500 | mg | ||
|
|
Vitamin B12 | 100 | mg | ||
|
|
Riboflavin | 100 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Filter-sterilize and store under a N2 atmosphere. | |||||