Transfection Optimization Strategy for the Nucleofector^TM Technology

In addition to data generated by Lonza, our comprehensive cell database also contains data on Nucleofection conditions that were shared by other users and which have provided sufficient results for their purposes. You may apply these conditions to your cell type of interest, but please be aware that a more comprehensive optimization using options A and B described above may provide better results for your specific cell clone or source. 

Using pmaxGFP^TM Positive Control for Transfection Optimization 

In addition to a complete optimization as described in option A above, we also provide a cell group-specific optimization protocol for a number of primary cell groups. Those so-called Basic Protocols refer to a pre-selected set of Primary Cell Nucleofector^TM Solutions (1 or 2) and a limited set of Nucleofector^TM Programs. Basic Protocols are available for the following cell groups and are suited for various mammalian species, e.g. human, mouse or rat. 

Performing a cell group-specific optimization using Basic Protocols

Performing a full optimization using Optimization Protocols

For each Nucleofector^TM Platform, we offer an optimization protocol for cell lines and for most platforms, we offer an optimization protocol for primary cells. For the Nucleofector^TM I/II/2b Device, we do not have an optimization protocol for primary cells due to the large number of different cell type specific solutions. Instead, we offer cell group-specific optimization protocols – refer to option B below.

The Cell Line Optimization Kit as well as the primary Cell Optimization Kit are working with a program matrix comprising 15 Nucleofection Programs for a first optimization. In most cases you will receive good results by applying this matrix only. However for maximum performance a fine tuning to reach higher efficiency or higher viability is recommended. The new kit concept enables this by offering an additional strip for fine tuning. Use these 16 reactions to further optimize your results. To do this:

• Select the best solution and the best program(s) from the first optimization round
  (for example DN-100)
• Use the table below to select alternative programs leading to higher viability
  (for example DH-100) or better efficiency (for example ER-100)

Note: Selecting pulses for higher viability will most probably led to a lower efficiency and pulses optimized for higher viability will lead to higher mortality. In case you can’t reach the desired performance, please contact our Scientific Support team for assistance.