Main sol. J1223
Internal Solution-ID: S5353
Equipment needed: Hungate tubes
Filter
Autoclave
Anaerobic workbench
| 1 Mix components, adjust pH to 7.5 and autoclave under a N2 gas atmosphere. After cooling, add the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized): | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 1.00 | g | 0.998 | 17.077 | |
| MgCl2 x 6 H2O | 0.40 | g | 0.399 | 1.964 | |
| CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.679 | |
| NH4Cl | 0.25 | g | 0.25 | 4.664 | |
| KH2PO4 | 0.20 | g | 0.2 | 1.467 | |
| KCl | 0.50 | g | 0.499 | 6.694 | |
| Na2SO4 | 4.00 | g | 3.992 | 28.105 | |
| Resazurin | 1.00 | mg | 1.0e-3 | 0.004 | |
| Distilled water | 988.00 | ml | - | - | |
| Trace element solution | 1.00 | ml | - | - | |
|
Vitamin solution
(*) |
1.00 | ml | - | - | |
|
Thiamine solution
(*) |
1.00 | ml | - | - | |
|
Vitamin B12 solution 0.05 g/L
(*) |
1.00 | ml | - | - | |
| Selenite-tungstate solution | 1.00 | ml | - | - | |
|
L-Sodium lactate
(1.0 M) |
7.30 | ml | - | - | |
| 2 Readjust pH to 7.5, if necessary. Separately autoclave and dry iron metal (grain). Aseptically distribute the medium and iron metal into culture vessels (e.g., 10 ml medium and 100 mg iron in Hungate tubes) under a N2 gas stream and seal with butyl rubber stoppers. Prior to use, add per liter the following solution: | |||||
|
Na2S x 9 H2O
(5%) |
2.00 | ml | - | - | |
Media with this solution
| J1223 | WIDDEL FRESHWATER MEDIUM WITH LACTATE |