621 .
HYL MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Final pH:
8.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HEPES | 4.76 | g | ||
|
|
NaCl | 20.00 | g | ||
|
|
Na2SO4 | 4.00 | g | ||
|
|
MgCl2 x 6 H2O | 4.30 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KCl | 0.34 | g | ||
|
|
CaCl2 x 2 H2O | 0.14 | g | ||
|
|
K2HPO4 | 0.14 | g | ||
|
|
Yeast extract | 1.00 | g | ||
|
|
Sodium lactate
(50%) |
4.50 | ml | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Autoclave and cool down to room temperature while flushing with N2. Then add the following solutions from sterile anaerobic stocks: | |||||
|
|
Trace elements | 1.00 | ml | ||
|
|
Fe(NH4)2(SO4)2 x 6 H2O
(in 0.037% HCl, 0.2%) |
1.00 | ml | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
Na2S x 9 H2O
(0.5 M) |
4.00 | ml | ||
| 2 Adjust pH to 7.0. | |||||
Trace elements #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 29.00 | g | ||
|
|
MnSO4 x n H2O | 3.26 | g | ||
|
|
CoCl2 x 6 H2O | 1.80 | g | ||
|
|
ZnSO4 x 7 H2O | 1.00 | g | ||
|
|
NiSO4 x 6 H2O | 0.11 | g | ||
|
|
CuSO4 x 5 H2O | 0.10 | g | ||
|
|
H3BO3 | 0.10 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.10 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.10 | g | ||
|
|
Na2WO4 x 2 H2O | 0.10 | g | ||
|
|
Na2SeO3 | 0.05 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA x 2Na and adjust pH to 8.0 with KOH. Then add the minerals. | |||||
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 |