Eppendorf Eporator - Manual (Page 14)

Operation
Eppendorf Eporator®
English (EN)
14
Abb. 5-2:Display
Fig. 5-2: Display
5.2 Recommendations for sample preparation
Independent of the device, the success of an electroporation is influenced by a variety of
factors:
• Quality and concentration of the inserted DNA
• Quality and concentration of the cells
• Resuspension medium of the DNA and cells
5.2.1 DNA preparation
• DNA quality: In order to obtain a high transformation efficiency, the DNA solution
should be pure and free of salts.
• Buffer: DNA dissolved in TE buffer is acceptable if the DNA was dissolved in approx.
ten times the quantity of electrocompetent cells.
• Salt concentration: DNA from enzyme reactions (e.g. ligation) can be immediately
used for electroporation if the salt concentration is under 5 M. If the ionic strength of
the reaction combination is too high, it can be reduced via dilution or ethanol
precipitation. After ethanol precipitation, the DNA can be resuspended in sterile,
demineralized water or TE buffer.
• Incubation: Before electroporation, do not incubate the DNA with the cell suspension
too long. Generally, the DNA should be added to the cells one minute before
electroporation and the solution should be incubated at 0 °C. Long incubation times
can lead to DNA degradation due to the DNases in the cell suspension.
1 Actual voltage value (in V)
2 Actual discharge time (in ms)
3 Voltage symbol
The voltage symbol appears after
electroporation and disappears after the
cuvette carrier has been removed.
4 Cuvette symbol
The cuvette symbol appears when a
cuvette is inserted.
5 Set voltage (in V)
1 2
3 4 5