4 electroporation protocol – Eppendorf Multiporator - Electroporation User Manual
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Table 2
Minimum pulse voltages at which the cell membranes may be permeated, in relation to the diameter of the cell after
incubation for 10 to15 minutes in the electroporation buffer, the electroporation temperature and the gap width of the
cuvette. Depending on the cell line, the optimal pulse voltage for the electroporation experiment may be between two
and five times higher.
* The maximum voltage which can be applied with the eukaryotic module of the Multiporator
®
is 1,200 V.
4.1.5 Preparing the DNA
The DNA should be ultra-pure (A260/A280
≥
1.8 ) and endotoxin-free. Following the last purification step, it should be
resuspended directly in bidistilled water and not in TE buffer.
4.1.6 Selecting the temperature
Electroporation may be carried out at room temperature or at 4 °C (see Sec. 3.6). Experiments to establish new protocols
normally take place at room temperature.
However, if electroporation takes place at 4 °C in order to achieve higher transfection rates, the cells should not remain
on ice for more than two minutes before being incubated at 37 °C.
Diameter of cell
[µm]
Voltage
2-mm cuvette
room temp.
Voltage
4-mm cuvette
room temp.
Voltage
2-mm cuvette
4 °C
Voltage
4-mm cuvette
4 °C
5
530
1100
1100*
*
10
270
540
540
1100
15
180
360
360
710
20
130
260
260
530
25
110
220
220
430
30
90
180
180
360
35
80
160
160
310
40
70
140
140
270
45
60
120
120
240
50
50
100
100
210
60
40
80
80
160
80
30
60
60
120
4 Electroporation protocol
4 Electroporation protocol
Multipor_Appli_E_poration_en.fm Seite 40 Montag, 30. Januar 2006 2:17 14