Eppendorf Eporator - Manual (Page 18)

Operation
Eppendorf Eporator®
English (EN)
18
5.3.3 Electroporation
5.4 Regeneration of the cells
Example for the bacterium E. coli:
1. After the electroporation, immediately place about 1 mL fresh medium (without
selection chemicals) on the cells. A rich medium is best suited for this, e.g. the SOC
medium for E. coli.
2. Carefully resuspend cells and transfer them to a new tube.
3. Incubate cells at optimal growth temperature (e.g. 37 °C for E. coli) for one hour at
light vibration (e.g. with the Eppendorf Thermomixer comfort).
5.5 Determination of the transformation efficiency
After the recovery period, the cells should be plated with a selection medium.
To determine the efficiency, streak different cell concentrations and use this information
to calculate the number of transformers/μg DNA.
1. Set a voltage between 200 V and 2 500 V using the arrow keys.
After switching on the device, the last set voltage is always displayed. The
most frequently used voltages can be saved and accessed using the program
keys (see Programs on p. 19).
2. Press the Start key to start the electroporation process.
• Charge and a progress bar appear in the display during loading.
• A signal tone sounds after the discharge.
• After the electroporation, the actual voltage (act), the discharge time of the
performed electroporation, and a voltage symbol appear in the display.
3. Remove cuvette holder from the device.
The cuvette symbol and voltage symbol disappear.
4. Remove electroporation cuvette from the cuvette holder and carefully
transfer the sample to the corresponding medium without bubbles.
Start