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Can I freeze the 96-well Nucleocuvette® Plates?

Yes. Freezing of the plates at -20°C does not alter their properties.

What does the message "weak" mean on the display of the Nucleofector® IIN, IIS or 2b after Nucleofection®?

This message means that the system cut off the pulse before it was completed (usually this is because the conductivity of the solution is at the low end of the normal range). The pulse was applied; it is just going to be a little weaker than...

Generation of a stable cell line: A positive control plasmid works, but I cannot obtain stable clones with my construct of interest. What could be the reason?

One reason could be that your gene product is toxic to the cells. We recommend testing the construct for expression of the gene of interest by transient Nucleofection® of your cells. If the proportion of expressing cells drops between 4 and 48 hours...

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...

Where can I find the meaning of the 4D-Nucleofector® System Error Codes?

In the back of the respective manuals, there is a chart giving a brief description of the error codes. If more information is needed or an error code is recurrent, it is best to contact Scientific Support for advice to determine if there is a...

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.

In stable cell line generation, I have a very high transfection efficiency, but most of my cells die during selection. Is this to be expected?

This is the normal pattern you should expect to see. Only a small proportion (1/10,000 to 1/100) of all transfected cells will integrate the transfected DNA into their genome and become stable transfectants. The remaining cells lose the transfected...

Can I use the Nucleofector® 2b Solutions and Optimized Protocols with the 4D-Nucleofector® X-Unit or the 96-well Shuttle® System?

No. The 4D-Nucleofector and 96-well Shuttle® System uses specifically optimized electrical parameters and solutions. Five different 4D-Nucleofector or 96-well Shuttle® Solutions are available for primary cells. For cell lines, three...

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.

At which time point after Nucleofection® should I start selection of stable integration?

Antibiotics for selection should be added 24-48 h after Nucleofection®.
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