Pay careful attention to centrifugation speeds prior to Nucleofection®; be sure not to centrifuge the cells at higher than 90xg.

Any trypsinization should use the minimal amount of reagent and time necessary in order to minimize stress to the cells, since the Nucleofection® will stress the cells as well. Neutralizing the trypsin with serum-containing media or trypsin inhibitor is imperative.

Post Nucleofection®, add the recommended amount of pre-warmed medium to the cuvette and gently transfer the cells with the provided pipettes without pipetting up and down (the key is to be as gentle as possible, especially at this step).

For other tips and suggestions, please watch our webinar on Tips for successful transfection experiments | Lonza