1033 .
ST65 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| 1 Mix components, except sulfur, bring to a boil and cool down under a CO2 (100%) gas stream. While gassing, add the following components to the medium: | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 18.000 | g | ||
|
|
MgCl2 x 6 H2O | 4.330 | g | ||
|
|
NH4Cl | 0.330 | g | ||
|
|
KCl | 0.330 | g | ||
|
|
KH2PO4 | 0.165 | g | ||
|
|
CaCl2 x 2 H2O | 0.033 | g | ||
|
|
Trace elements solution | 10.000 | ml | ||
|
|
Sulfur
(powder) |
5.000 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
|
|
Trace vitamins | 10.000 | ml | ||
|
|
NaHCO3 | 2.000 | g | ||
| 2 Bubble CO2 for 3 - 4 min and check pH to be 6.5 - 6.8. Readjust pH to the range if necessary. Autoclave under a CO2 gas atmosphere. Add the following solution to the medium (separately autoclaved under N2): | |||||
|
|
KNO3
(1 M) |
10.000 | ml | ||
| 3 Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 10 ml of the medium and 0.05 g sulfur in Hungate tubes) under a CO2 stream and seal with butyl rubber stoppers. | |||||
Trace elements solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Fe(NH4)2(SO4)2 x 6 H2O | 78.40 | mg |
|
|
CoCl2 x 6 H2O | 23.80 | mg |
|
|
NiCl2 x 6 H2O | 16.60 | mg |
|
|
Na2MoO4 x 2 H2O | 2.42 | mg |
|
|
Na2WO4 x 2 H2O | 5.49 | mg |
|
|
ZnSO4 x 7 H2O | 14.40 | mg |
|
|
CuCl2 x 2 H2O | 0.17 | mg |
|
|
Na2SeO4 | 9.45 | mg |
|
|
H3BO3 | 0.62 | mg |
|
|
MnCl2 x 4 H2O | 19.80 | mg |
|
|
SrSO4 | 0.92 | mg |
|
|
CrK(SO4)2 x 12 H2O | 0.50 | mg |
|
|
AlCl3 x 6 H2O | 2.41 | mg |
|
|
BaCl2 | 2.08 | mg |
|
|
Na2SiO3 x 9 H2O | 14.20 | mg |
|
|
KBr | 11.90 | mg |
|
|
KI | 16.60 | mg |
|
|
Na2SO4 | 71.00 | mg |
|
|
Distilled water | 1000.00 | ml |
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 |