1177 .
SEDIMENTISPHAERA L21 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 120.0 | g | ||
|
|
MgCl2 x 6 H2O | 6.0 | g | ||
|
|
KCl | 1.5 | g | ||
|
|
Na2SO4 | 1.0 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
CaCl2 x 2 H2O | 0.4 | g | ||
|
|
K2HPO4 | 0.4 | g | ||
|
|
Trace mineral solution | 10.0 | ml | ||
|
|
Resazurin | 0.5 | mg | ||
|
|
Distilled water | 950.0 | ml | ||
| 1 Mix components thoroughly, bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream and autoclave under the same gas mixture. After cooling, add per liter the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
Glucose
(10%) |
10.0 | ml | ||
|
|
Trace vitamins
(*) |
10.0 | ml | ||
|
|
NaHCO3
(*, 8%) |
30.0 | ml | ||
| 2 Aseptically and anaerobically, dispense the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a N2-CO2 (4:1, v/v) gas stream and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N2 atmosphere): | |||||
|
|
L-Cysteine HCl x H2O
(5%) |
10.0 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
10.0 | ml | ||
| 3 Readjust pH to 7.5, if necessary. | |||||
Trace mineral solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Trace minerals | 1000.00 | ml |
|
|
NiCl2 x 6 H2O | 0.03 | g |
|
|
Na2SeO3 x 5 H2O | 0.30 | mg |
|
|
Na2WO4 x 2 H2O | 0.40 | mg |
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 |