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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...

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.

According to which standard is the Nucleofector® System certified?

The 4D-Nucleofector® Unit was tested by TÜV Rheinland Product Safety GmbH and found to be in compliance with the following standards:• IEC 61010-1The 4D-Nucleofector® Unit was tested by TÜV Rheinland of North America Inc. and found to be in...

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...

Is it typical for the fluorescence observed from my tagged protein to appear less than the empty fluorescent vector?

Yes, this is a routine observation.Fluorescent fusion proteins can appear less bright than the fluorescent protein itself. This may be due to differences in protein folding of the fused protein.

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...

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.

In my Nucleofection® Experiment, how critical is the time point for analysis post transfection?

It is very critical. Waiting too long for analysis post transfection can lead to loss of peak expression. The optimal time for analysis is related to two factors: stability of protein being expressed and transfection method. For a short-lived protein...

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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