Optimization Protocol for DNA and siRNA,
Continued
24-well
Optimization
Protocol for
Adherent and
Suspension Cell
Lines—Day One,
Continued
5.
After electroporation, immediately remove the Neon
™
Pipette and transfer
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). Select the
best conditions and proceed to the next day’s experiment, page 27.
Continued on next page
25
