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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...

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.

Can mycoplasma grow in the absence of cells?

There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present. 

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.

Is it possible to store reconstituted MycoAlert®/MycoAlert® PLUS Reagent and Substrate?

For optimal assay conditions, reconstituted reagent and substrate should be used fresh within 2 hours after reconstitution.During this time they can be kept at room temperature.For the MycoAlert® Kit: Unused, reconstituted components can be aliquoted...

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...

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.

Do we sell the Nucleofector® Solution (or any other component of the Nucleofector® Kit) separately?

We don't sell the individual components of the kits separately because our product line is a kit concept line to ensure the use of certified products with quality controlled lot numbers that guarantee optimal results.

How much sample do you need for accurate results with MycoAlert® Assay? Do you need cells or just supernatant?

The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.
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