Optimization Protocol for DNA and siRNA,
Continued
Optimization Protocol—Day Two,
Continued
Results
Sample Well
no. Pulse
Voltage
Pulse
Width
Pulse
no.
Transfection Efficiency Cell Viability
1 A1 1450 20 1
2 A2 1475 20 1
3 A3 1500 20
1
4 A4 1525 20 1
5 A5 1550 20 1
6 A5 1575 20 1
7 B1 1375 30 1
8 B2 1400 30
1
9 B3 1425 30 1
10 B4 1450 30 1
11 B5 1475 30 1
12 B6 1500 30 1
13
C1
Control containing DNA but no electroporation pulse.
5.
After electroporation, immediately remove the Neon
™
Pipette and transfer
the samples from the 10 μL Neon
™
Tip into prewarmed 0.5 mL culture
medium.
For 100 μL Neon
™
Tip, dilute samples 10-fold in 900 μL medium and
transfer 100 μL of the sample to 0.4 mL prewarmed culture medium.
6.
Repeat Steps 3–5 for the remaining samples.
7.
Gently rock the plate to assure even distribution of the cells. Incubate the
plate at 37°C in a humidified CO
2
incubator.
8.
Assay samples to determine the transfection efficiency (e.g., fluorescence
microscopy or functional assay) or gene knockdown (for siRNA).
9.
Select the best conditions and proceed to the next day’s experiment, next
page.
Continued on next page
28
