Cell lines stably expressing a target gene are often used for drug screening. However the number of stably expression clones resulting from lipofection experiments is often low, especially for cells which are difficult-to-transfect. Due to its high efficiencies for transient transfections, Lonza’s Nucleofector™ Technology is also well suited for the generation of stable expressing clones in difficult-to-transfect cell types.

 

Efficient non-viral transfection


  • Up to 90% transfection efficiency for primary cells and difficult-to-transfect cancer cell lines, including blood, breast and prostate cancer
  • Combined with high cell viability
  • Outperforms lipofection for integration rates

 

Proven technology

  • Minimal optimization effort due to availability of more than 100 ready-to-use Nucleofection™ Protocols
  • More than 800 cancer-related publications

4D Nucleofector Device 

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Higher integration rates in difficult-to-transfect cell lines using Nucleofection™.
Jurkat cells were transfected with linearized or circular DNA using either Nucleofection™ (2 µg DNA) or competitor lipofection reagent L (0.7 µg DNA) according to manufacturers instructions. 24 hours after transfection cells were G418 was added for selection of stably transfected cells. 30 days after plating cells were analyzed for clonal outgrowth (Integration frequency = number of resistant clones per number of living cells seeded). Due to toxic effects lipofection with 10 µg circular DNA could not be performed.

 

Basics about Nucleofector™ Technology


Nucleofection™ is a technology based on the momentary creation of small pores in cell membranes by applying an electrical pulse. The comprehensive way in which Nucleofector™ Programs and cell type-specific solutions are developed enables nucleic acid substrates delivery not only to the cytoplasm, but also through the nuclear membrane and into the nucleus. This allows for high transfection efficiencies up to 99% and makes the transfection success independent from any cell proliferation.

Developed in 1998, the Nucleofector™ Technology was introduced to the research market in 2001 as the first efficient non-viral transfection method for primary cells and hard-totransfect cell lines. With the Nucleofector™ Technology primary cells and stem cells, as well as cell lines, can be consistently transfected at high efficiency. Since then the technology has evolved through constant innovation.

   

 

 

 



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