Tested Electroporation Conditions Using the siPORTer™-96
Cell Type
Voltage
(Volts)
Pulse Length (microseconds)
Number of Pulses
Time Between Pulses (seconds)
Control siRNA Used*
Number of Cells
% mRNA Remaining*
% Viability*
NHDF-neo (normal human dermal fibroblasts-neonatal)
600
400
1
-
GAPDH
25000
3.8
90
RPTEC (human renal proximal tubule cells)
350
350
1
-
GAPDH
25000
15
92
hMSC (human mesenchymal stem cells)
800
200
1
-
GAPDH
25000
7.5
90
HUVEC (human umbilical vein endothelial cells)
200
200
1 to 2
0.1
GAPDH
25000
28
90
MEF (mouse embryo fibroblsts)
500
200
1
-
GAPDH
25000
32
88
RAW264.7 (mouse macrophage)
300
400
1
-
KIF11 (Eg5)
25000
18
84
RMSC (rhesus monkey stem cells)
300
250
1 to 2
0.1
GAPDH
25000
7.5
87
Jurkat (acute T-cell leukemia)
200
250-350
1 to 2
0.1
GAPDH
200000
28
75**
T cells (human, primary)
450
400
1
-
GAPDH
200000
15
67**
* Experiments were performed by electroporating eight identical samples simultaneously with the siPORTer-96 using the above pulse conditions. Note that the voltage value listed above represents the voltage setting on the Gene Pulser XCell™ power supply, not the voltage across each sample. One µg of the indicated control siRNA or Silencer® Negative Control siRNA was electroporated into the indicated number of cells in siPORT™ siRNA Electroporation Buffer at a final volume of 45 µl. 24 hours post-electroporation, mRNA levels were monitored by real-time RT-PCR, and cell viability was monitored using the Guava ViaCount assay. '% mRNA Remaining' was calculated relative to cells electroporated with Silencer Negative Control #1 siRNA. Viability is expressed relative to the total population after electroporation.
**Viability of Jurkat cells prior to electroporation = 85%; Viability of primary T-cells prior to electroporation = 88%.