Optimization Protocol for DNA and siRNA,
Continued
Optional: Optimization Protocol—Day Three,
Continued
Results
Sample Well
no. Pulse
Voltage
Pulse
Width
Pulse
no.
Transfection Efficiency Cell Viability
1 A1 1450 10 2
2 A2 1475 10 2
3 A3 1500 10
2
4 A4 1525 10 2
5 A5 1550 10 2
6 A6 1575 10 2
7 B1 1375 10 3
8 B2 1400 10
3
9 B3 1425 10 3
10 B4 1450 10 3
11 B5 1475 10 3
12 B6 1500 10 3
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 and incubate the plate.
7.
Assay samples to determine the transfection efficiency (e.g., fluorescence
microscopy or functional assay) or gene knockdown (for siRNA).
8.
Select the best conditions and save these parameters into the database for
your cell type.
30
