Main sol. 1012c
Internal Solution-ID: S6442
Equipment needed: Autoclave
Petri dishes
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
HEPES buffer
(SIGMA H3375) |
0.72 | g | - | - | |
| CaCl2 x 2 H2O | 0.15 | g | - | - | |
| MgCl2 x 6 H2O | 0.02 | g | - | - | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Grow prey bacteria on agar plates using the medium indicated below. Use one plate of a 24-hour-old culture and wash off grown cells in 5 ml sterile water. | |||||
| 2 Prepare HEPES buffer, adjust pH to 7.5, and sterilize by filtration. After cooling to room temperature add 5 ml cell suspension of the host organism to 50 ml HEPES buffer and mix. The prey medium suspension should contain approx. 107 to 109 cells. Inoculate the complete medium immediately after preparation and incubate the suspension with shaking until a decrease in turbidity is visible. | |||||