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6401 results sorted by

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.

Are there any recommendations to avoid excessive mortality during T cell Nucleofection®?

Try to keep the DNA amount for Nucleofection® quite low (e.g. 1 µg plasmid DNA per 100 µl reaction) since higher DNA amounts might cause increasing toxicity and mortality. It is also important to use highly purified DNA, e.g. by using Endofree™...

Following Nucleofection®, I cannot obtain stable clones from single cells. What could be the reason?

Some cell lines need certain cell densities to proliferate. For those cells you can try using conditioned medium for setting up limiting dilution or culturing single cells together with feeder cells.

Can I freeze the 96-well Nucleocuvette® Plates?

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

What antibiotic concentration should I use for selecting stable transfectants?

We highly recommend to perform a titration for G418 in a range of 0,1 mg/ml to 1,5 mg/ml; in serum-free culture expand range down to 20 µg/ml).Choose the concentration which is 0,1 or 0,2 mg/ml above the one which shows complete cell death as...

How much DNA should I use in my Nucleofection® Reaction to obtain stable clones?

You should use the amount of DNA recommended in the optimized protocol for transient transfection. If obtaining a very high number of clones is important for your experiments, you can try increasing the amount of DNA. For further tips see our...

How can I increase cell viability of Dendritic cells after Nucleofection®?

DNA amount and quality are very critical for Nucleofection® of Dendritic Cells (recommended 0.5-1 µg; maximal 2 µg). LPS (lipopolysaccharides) have a strong negative effect on cell viability. Please make sure that all components you use for dendritic...

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...

I want to stimulate Human T cells post Nucleofection®. Are there any precautions to keep in mind?

After Nucleofection®, wait at least four hours before stimulation. Stimulating immediately after Nucleofection® may lead to increased cell mortality and poor activation of cells. For more details on the stimulation of Human T cells post...

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...
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