Main sol. 78
Internal Solution-ID: S123
This solution is used in 5 media and was first defined in medium 78.
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Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Ground beef
(fat free) |
500.0 | g | 500 | - | |
| Distilled water | 1000.0 | ml | - | - | |
|
NaOH
(1 N) |
25.0 | ml | - | - | |
| 1 Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add water to a final volume of 1000 ml, and then add: | |||||
| Casitone | 30.0 | g | 30 | - | |
| Yeast extract | 5.0 | g | 5 | - | |
| K2HPO4 | 5.0 | g | 5 | 28.707 | |
| Resazurin | 1.0 | mg | 1.0e-3 | 0.004 | |
| 2 To make medium anoxic bring it to a boil, cool under 100% N2 gas atmosphere, add 0.5 g/l L-cysteine hydrochloride and adjust pH to 7.0. Dispense under 100% N2 gas atmosphere by filling 7 ml medium into anoxic Hungate-type tubes (for strains demanding meat particles put those first into the tube (use 1 part meat particles to 4 or 5 parts fluid)). Autoclave at 121°C for 30 min. For agar slants use 15.0 g agar per 1000.0 ml medium. | |||||
| L-Cysteine HCl | 0.5 | g | 0.5 | 3.172 | |
| Agar , for solid medium | 15.0 | g | 15 | 44.599 | |
| 3 In some cases (as indicated in the catalogue) the addition of Haemin and Vitamin K1 or Vitamin K3 is necessary. Add to 1000 ml of medium after autoclaving: | |||||
|
Haemin solution
optional |
10.0 | ml | - | - | |
|
Vitamin K1 solution
, alternative
optional |
10.0 | ml | - | - | |
|
Vitamin K3 solution
, alternative
optional |
10.0 | ml | - | - | |