665 .
FRESHWATER IRON GEL MEDIUM
What is this page?
Equipment needed:
Autoclave
Centrifuge
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Poorly crystalline iron(III) oxide | 100.00 | mM | ||
|
|
NaHCO3 | 2.50 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
NaH2PO4 x H2O | 0.60 | g | ||
|
|
KCl | 0.10 | g | ||
|
|
Na2SeO4
(1 mM) |
1.00 | ml | ||
|
|
Vitamin solution | 10.00 | ml | ||
|
|
Trace mineral solution | 10.00 | ml | ||
| 1 Add components to distilled water, bring volume to 1.0 L and adjust pH to 7.0. Distribute the medium into culture vessels under a N2-CO2 (4:1, v/v) gas mixture, sparge with the same gas mixture for 10 - 15 min, seal with butyl rubber stoppers and autoclave. Before inoculation, add the following sterile solutions (autoclaved and stored under a N2 gas atmosphere) to the medium (per 10 ml of medium): | |||||
|
|
FeCl2
(130 mM) |
0.10 | ml | ||
|
|
CaCl2 x 2 H2O
(0.4%) |
0.10 | ml | ||
|
|
Sodium acetate
(pH 7.0, 2 M) |
0.10 | ml | ||
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20.0 | 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 |
|
|
DL-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 |
Poorly crystalline iron(III) oxide #
| 1 Dissolve 108 g FeCl3 in 1.0 L distilled water. Adjust pH to 7.0 with 10 N NaOH. Allow the suspension to sit for 30 min, then readjust pH to 7.0. Centrifuge the suspension at 5,000 x g for 20 min and pour off supernatant. Wash the precipitate with distilled water by resuspending in distilled water followed by the centrifugation. Repeat the washing at least 5 times, then, resuspend in 1.0 L distilled water. Use the Fe(III) suspension as 0.67 M solution. More precisely, check the concentration of Cl- to be less than 1 mM and determine the concentration of Fe(III) by using hydroxylamine as described by Lovley and Phillips (Appl. Environ. Microbiol. 54, 751-757, 1986). |
Trace mineral solution #
| 1 Use Trace minerals of Medium No. 151 supplemented with 0.025 g (final) Na2MoO4 x 2H2O, 0.024 g NiCl2 x 6H2O and 0.025 g Na2WO4 x 2H2O (per liter). | |||||
| 2 Comment: For strain JCM 12378, inoculate with a 10% inoculum. |