44 Life Technologies
™
| Alternative Transfection Method: Neon
®
Transfection System
Transfect within 10 minutes of cell resuspension in Resuspension Buffer R
• We recommend only resuspending sufficient cells for 6 repeats at
a time. If more than 6 repeats are desired, we suggest centrifuging
the total desired number of cells in the first PBS wash step, but
only performing the second PBS wash and final cell resuspension
using a maximum of 5.0 x 10
7
viable cells at a time.
Use a new Neon
®
Tube for each new plasmid DNA or cell type
being transfected.
Change the Neon
®
Tube after 10 uses.
Use a new Neon
®
Tip for each new plasmid DNA or cell type
being transfected.
Change the Neon
®
Tip after two uses.
Ensure that there are absolutely no air bubbles in the Neon
®
Tip before
proceeding with electroporation (Figure 7). It may take a few attempts
to properly fill the Neon
®
Tip.
Don't activate the current if there are air bubbles in the Neon
®
Tip
(Figure 7).
Alternative Transfection Method: Neon
®
Transfection System
Don't:
Do:
