1840 .
Halorutilales haloarchaeon medium
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Equipment needed:
Autoclave
Final pH:
6.8
Description:
used for cultation of lithotrophic thiosulfate-oxidizing Halorutilales haloarchaeon strain col2-1 (DSM 117793)
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 240.00 | g | ||
|
|
K2HPO4 | 2.50 | g | ||
|
|
KCl | 5.00 | g | ||
|
|
NH4Cl | 0.20 | g | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Distilled water | 999.00 | ml | ||
| 1 Adjust pH to 6.8 with 0.5 M KH2PO4. | |||||
| 2 Dispense into serum bottles (30 ml medium in 100 ml serum bottles, OR 15 ml in 50 serum bottles). | |||||
| 3 add 20% CO2 to gas phase (in serum bottles) | |||||
| 4 Autoclave at 121 °C for 20 min. | |||||
| 5 Before inoculation add from sterile stock solutions per liter: | |||||
|
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Vitamine mixture | 0.50 | ml/l | ||
|
|
MgSO4 x 7 H2O
(1M in water) |
0.08 | ml/l | ||
|
|
alkaline trace metal solution | 0.50 | ml/l | ||
|
|
Na-thiosulfate
(2M in water) |
10.00 | ml/l | ||
|
|
NaHCO3
(8% in water) |
10.50 | ml/L | ||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
|
FeSO4 x 7 H2O | 2.00 | g | ||
|
|
ZnSO4 | 0.10 | g | ||
|
|
MnCl2 | 0.03 | g | ||
|
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H3BO3 | 0.30 | g | ||
|
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CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 5 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.02 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 3.0-4.0 with HCl. | |||||
| 2 Autoclave at 121 °C for 15 min. | |||||
| 3 Note: autoclave in closed bottles | |||||
Vitamine mixture #
400
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Acidic thiamine / Ca pantothenate solution | 100 | ml | ||
|
|
Basic vitamin solution | 100 | ml | ||
|
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Neutral folic acid / riboflavin solution | 100 | ml | ||
|
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Neutral B12 solution 0.005 g/L | 100 | ml | ||
| 1 Sterilize by filtration. | |||||
| 2 Mix the four solutions in the ratio 1:1:1:1. | |||||
Acidic thiamine / Ca pantothenate solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine | 5 | mg |
|
|
Calcium pantothenate | 5 | mg |
|
|
Distilled water
(acidified to pH 3.0 with HCl) |
100 | ml |
Basic vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2 | mg |
|
|
PABA
(p-aminobenzoic acid) |
5 | mg |
|
|
Nicotinic acid | 5 | mg |
|
|
Pyridoxine | 10 | mg |
|
|
NaOH
(0.1N) |
100 | ml |
Neutral folic acid / riboflavin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Folic acid | 2 | mg |
|
|
Riboflavin | 5 | mg |
|
|
Distilled water | 100 | ml |
alkaline trace metal solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.040 | g/l |
|
|
Na2SeO3 x 5 H2O | 0.026 | g/l |
|
|
Na2WO4 x 2 H2O | 0.030 | g/l |
|
|
Distilled water | 1000.000 | ml |