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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...

Can calcium influx issues affect viability or differentiation in Stem Cells after Nucleofection?

When cells are transfected by Nucleofection™, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection™ in medias that contain...

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...

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...

What are the requirements for direct Nucleofection of mRNA for protein expression?

The mRNA should be capped and polyadenylated. The conditions of Nucleofection™ will be the same as for DNA with the particular cell type, i.e. follow the same protocol and use the same program, except one will likely need to add a much higher amount...

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.

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....

Is there a reason why in our Nucleofector Protocols for Primary Human Chondrocytes we include FCS in the pronase and collagenase solutions for preparation of chondrocytes?

Yes, even though this may sound odd since serum is generally known to inactivte proteases. We recommend it in this case, however, because the incubation times are quite long and the cells are stressed by the procedure. The amounts of enzymes should...

Does the GFP sequence in your pmaxGFP control plasmid have a stop codon?

Yes. The coding sequence ends with a stop codon (TGA/UGA).

Do you have a protocol for the Nucleofection of Plasmodium berghei?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector™ Solution for your...
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