210 .
HALOANAEROBIUM MEDIUM
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Equipment needed:
Anaerobic workbench
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Final gas composition:
100 % N2
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Final pH:
9.0
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 130.0 | g | ||
|
|
MgSO4 x 7 H2O | 8.8 | g | ||
|
|
KCl | 1.0 | g | ||
|
|
Yeast extract | 10.0 | g | ||
|
|
Trypticase
(BBL) |
10.0 | g | ||
|
|
Wolin's vitamin solution | 10.0 | ml | ||
|
|
Trace element solution | 10.0 | ml | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Prepare the medium anaerobically under 100% N2. After autoclaving add from sterile solutions: | |||||
|
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Thioglycolate-ascorbate solution | 25.0 | ml | ||
|
|
Glucose
(10 % w/v) |
25.0 | ml | ||
|
|
NaOH
(2 N) |
10.0 | ml | ||
| 2 Final pH: 9.0. | |||||
Thioglycolate-ascorbate solution #
25
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na-thioglycolate | 0.5 | g | ||
|
|
Na-ascorbate | 0.5 | g | ||
|
|
Distilled water | 25.0 | ml | ||
| 1 Filter sterilize. | |||||
Wolin's vitamin solution #
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 D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid
(NTA) |
12.80 | g | ||
|
|
FeCl2 x 4 H2O | 0.20 | g | ||
|
|
MnCl2 x 4 H2O | 0.10 | g | ||
|
|
CoCl2 x 6 H2O | 0.17 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnCl2 | 0.10 | g | ||
|
|
CuCl2 | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.03 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml. | |||||