1101 .
HALOLACTIBACILLUS 9G MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
9.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 10.0 | g | ||
|
|
KCl | 0.2 | g | ||
|
|
K2HPO4 | 0.2 | g | ||
|
|
MgCl2 x 6 H2O | 0.1 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
Yeast extract | 0.1 | g | ||
|
|
Trace vitamins | 2.0 | ml | ||
|
|
Trace minerals solution | 1.0 | ml | ||
|
|
Resazurin | 0.5 | mg | ||
|
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Distilled water | 680.0 | ml | ||
| 1 Mix components thoroughly and autoclave under a N2 atmosphere in a bottle capped with a butyl rubber stopper. After cooling, add the following solution to the medium: | |||||
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Alkaline solution | 300.0 | ml | ||
|
|
Glucose
(autoclaved, 1.0 M) |
20.0 | ml | ||
| 2 Anaerobically and aseptically distribute the medium into culture vessels (e.g., 10 ml medium in Hungate tubes) under a N2 gas atmosphere and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution (autoclaved and stored under a N2 gas atmosphere): | |||||
|
|
Na2S x 9 H2O
(5%) |
8.0 | ml | ||
| 3 Adjust pH to 9.0, if necessary. | |||||
Alkaline solution #
300
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2CO3 | 2.76 | g | ||
|
|
NaHCO3 | 24.00 | g | ||
|
|
Distilled water | 300.00 | ml | ||
| 1 Mix components thoroughly and filter-sterilize. | |||||
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 |
Trace minerals solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.500 | g | ||
|
|
Fe(NH4)2(SO4)2 x 6 H2O | 0.200 | g | ||
|
|
Na2SeO3 | 0.100 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.100 | g | ||
|
|
Na2WO4 x 2 H2O | 1.000 | g | ||
|
|
MnSO4 x n H2O | 0.100 | g | ||
|
|
ZnSO4 x 7 H2O | 0.100 | g | ||
|
|
NiCl2 x 6 H2O | 0.025 | g | ||
|
|
CuSO4 x 5 H2O | 0.010 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. | |||||
| 2 Adjust final pH to 7.0. | |||||