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Can I co-transfect oligonucleotides and plasmids with the Nucleofector® Technology?

Yes. As the same protocol applies for any nucleic acid substrate (vectors or oligonucleotides) you can easily perform co-transfections either for transfection control or rescue experiments.

Which pipette tips are compatible with the 96-well Shuttle® Nucleocuvette® Plates?

We recommend using special tips, e.g. epT.I.P.S™ (Eppendorf, Hamburg, Germany) or TallTips™ (Matrix Technologies, Hudson, NH, USA). You can contact our Scientific Support Team for a list of compatible tips, including tips for pipetting robots.

Can I integrate the 96-well Shuttle® Device into a liquid handling system?

Yes. The 96-well Shuttle® Device is designed in such a way that it is addressable by robot grippers. Furthermore, the software contains the required interface to be connected with the LHS software. However, the practical implementation for each...

Why does Lonza recommend using non-ventilated caps when culturing insect cells?

The reason is that insect cells are often incubated in very simple incubators that are not humidified (and without CO2 at 24°C to 28°C). Insect media typically do not need CO2 for buffering pH, so gas exchange is not required. To avoid evaporation of...

Can I use a Nucleofector® I or Nucleofector® 2b Device together with the 96-well Shuttle® Device?

No, the 96-well Shuttle® only works in conjunction with the Nucleofector® IIs Device or our 4D-Nucleofector® Device.

How long does it take to obtain stable transfectants?

Depending on individual cell type and doubling rate, selection of stable transfectants will take between 7 and 28 days. Expansion and characterization of single cell clones will take several weeks in addition.

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...

What selection markers can I use for the generation of stable cell lines?

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.

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.

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

The culture medium should be changed every 2-3 days during the selection process.This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.
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