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Tested Electroporation Conditions for Various Cell Types

Cell Type

Voltage
(volts)

Pulse Length
(microseconds)

Number of Pulses

Time Between Pulses (seconds)

Number of Cells

% mRNA Remaining*

% Viability*

BAEC (bovine aortic endothelial cells)

250 V

160 µS

1-2

0.1 s

75,000

20
85

BAMEC (bovine adrenal microvascular endothelial cells)

300 V

200 µS

1

-

75,000

32
85

BAVSMC (bovine aortic vascular smooth muscle cells)

300 V

100 µS

1-2

0.1 s

75,000

30
70

BLMVEC (bovine lung microvascular endothelial cells)

300 V

140 µS

1-2

0.1 s

75,000

32
75

hMSC (human mesenchymal stem cells)

700 V

90 µS

2

5 s

75,000

1
95

HUVEC (normal human umbilical vein endothelial cells)

250 V

150 µS

1

-

75,000

9
70

K562 (human lymphoblasts)

300-350 V

130 µS

1-2

0.1 s

150,000

23
75

MEF (mouse embryo fibroblasts)

350 V

150 µS

2

0.1 s

75,000

13
85

NHA (normal human astrocytes)

400 V

100 µS

2-3

0.1 s

75,000

7
85

NHDF-neo (normal human dermal fibroblasts-neonatal)

900 V

70 µS

2

5 s

75,000

2
85

PC-12 (rat pheochromocytoma)

450 V

200 µS

1

-

75,000

12
80

Rat astrocytes

300 V

90 µS

2-3

0.1 s

75,000

14
85

RMSC (rhesus monkey stem cells)

450 V

120 µS

2

0.1 s

75,000

11
85
Notes:

1. All electroporation reactions were performed with square wave type pulses.

2. Used 1.5 µg siRNA and 75 µl siRNA electroporation buffer per reaction.

3. Electroporations were performed in standard 1 mm electroporation cuvettes (Bio-Rad).  For cuvettes of other sizes, please adjust voltages accordingly.

4. % mRNA remaining is reported relative to samples electroporated with Silencer® Negative Control #1 siRNA. % viability is reported relative to non-treated cells.

5. These conditions are provided as a starting point for your convenience.  Even if you are using the same types of cells as listed here, for the best gene silencing results, we recommend that you try to optimize these parameters.


siPORT™ siRNA Electroporation Buffer Information

 
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