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What selection markers can I use for stable transfections?

The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.

What is the advantage of the 96-well Shuttle Device over common lipofection, especially in a high throughput framework?

Several primary cells of high research relevance can simply not be transfected at relevant efficiencies and viabilities using lipofection. The same holds true for many suspension cell lines which are often used for protein production. These difficult...

What are the contents of a 96-well Shuttle Kit?

As the system will most likely be used for high-throughput approaches we provide plates which are pre-equipped with 96-well Nucleocuvette™ modules (2x8). In addition to that, the 96-well Shuttle solution, the supplement, and the pmaxGFP™ are also...


How often should I change the medium during selection of stable clones after Nucleofection?

Every 2-3 days. This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.

What is the advantage of the Nucleofector Technology over common electroporation (systems)?

High cell viability and high transfection efficiency with the Nucleofection™ Technology are the most prominent features. Nucleofection™, i.e., the transfer of DNA into the nucleus and not only the cytosol, is essential when working with non-dividing...

How many stable clones will I get from one transfection?

This depends on the respective cell type (transfection efficiency, viability and integration frequency). If it is important to you to know the frequency of stable integrants before carrying out your actual experiments, the frequency should be...

Is it possible to pre-plate substrates when using the 96-well Shuttle for Nucleofection?

Yes, pre-plating of substrates like siRNA and subsequent storage at -20°C is possible.

Following Nucleofection, I cannot obtain stable clones from single cells, what could be the reason?

Some cell lines need certain cell densities to proliferate. For those cells you can try using conditioned medium for setting up limiting dilution or culturing single cells together with feeder cells.

What kind of protocol you recommend for stimulation of human T cells?

For getting the results we present in our datasheet, we recommend the follwing strategy: Cells should be stimulated and cultivated for 3 days before Nucleofection, preferentially on plates coated with 1 µg/ml anti-CD3 and 2 µg/ml anti-CD28 antibody....
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