1004 .
FRESHWATER MEDIUM FOR SOIL AOA
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Equipment needed:
Autoclave
Filter
Final pH:
7.0
Select a strain (optional):
Basic FWM solution #
1017
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.0 | g | ||
|
|
KCl | 0.5 | g | ||
|
|
MgCl2 x 6 H2O | 0.4 | g | ||
|
|
KH2PO4 | 0.2 | g | ||
|
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Autoclave Basic FWM solution and aseptically add the following solutions (autoclaved or *filter-sterilized) in the given order: | |||||
|
|
Modified trace element mixture | 1.0 | ml | ||
|
|
Vitamin solution
(*) |
1.0 | ml | ||
|
|
FeNa-EDTA
(pH 7.0, *, 7.5 mM) |
1.0 | ml | ||
|
|
NaHCO3
(*, 1 M) |
2.0 | ml | ||
|
|
HEPES solution | 10.0 | ml | ||
|
|
NH4Cl
(1 M) |
1.0 | ml | ||
|
|
Sodium pyruvate
(*, 1 M) |
1.0 | ml | ||
| 2 Use glass or polystyrene containers for cultivation. Cultivate in the dark without shaking. | |||||
Modified trace element mixture #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(25%) |
12.5 | ml |
|
|
H3BO3 | 30.0 | mg |
|
|
MnCl2 x 4 H2O | 100.0 | mg |
|
|
CoCl2 x 6 H2O | 190.0 | mg |
|
|
NiCl2 x 6 H2O | 24.0 | mg |
|
|
CuCl2 x 2 H2O | 2.0 | mg |
|
|
ZnSO4 x 7 H2O | 144.0 | mg |
|
|
Na2MoO4 x 2 H2O | 36.0 | mg |
|
|
Distilled water | 987.0 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 20 | mg | ||
|
|
Folic acid | 20 | mg | ||
|
|
Pyridoxine hydrochloride | 100 | mg | ||
|
|
Thiamine HCl | 50 | mg | ||
|
|
Riboflavin | 50 | mg | ||
|
|
Nicotinic acid | 50 | mg | ||
|
|
DL-Calcium pantothenate | 50 | mg | ||
|
|
Vitamin B12 | 1 | mg | ||
|
|
p-Aminobenzoic acid | 50 | mg | ||
|
|
Choline chloride | 2 | g | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH to 7.0 with KOH. | |||||
HEPES solution #
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaOH | 12.0 | g | ||
|
|
HEPES
(free acid) |
119.2 | g | ||
| 1 Dissolve NaOH in 300 ml distilled water. While stirring vigorously, slowly add HEPES until completely dissolved. Fill up to 450 ml with distilled water. If 1 ml of this solution is added to 100ml of the medium (containing all additive but HEPES), pH should be around 7.6 at 30C. Otherwise, adjust pH of the HEPES solution with 10 N NaOH or conc. HCl. Finally fill up to 500 ml with distilled water. | |||||