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What is the optimal size of DNA that you recommend for Nucleofection?

We routinely use plasmids of 4-7 kb in our laboratories and plasmids up to approximately 20kb should not be a problem. Using plasmids larger than this will most likely result in lower transfection efficiency. Some preliminary results we have also...

How should I purify my DNA for Nucleofection of neurons?

The quality and the concentration of DNA used for Nucleofection™ in general plays a central role for the efficiency of gene transfer. We strongly recommend using endotoxin free prepared DNA. Endotoxin free Kits are available from several suppliers....

I've noticed that duration of the Nucleofector Program seems to change even when I haven't changed the program I am using. Why?

The duration time of a program is dependent on the number of the program. The duration is also dependent on how long the Nucleofector™ Device has been turned on as well as the number of Nucleofections you have done in the last couple of minutes....

What is the lowest amount of DNA which can be used for transfection? Do I need a DNA carrier for low DNA amounts?

The recommended amount of DNA per 100 µl reaction is 0.5µg-5 µg. No carrier is needed.

Can I use larger cuvettes for my Nucleofection Reaction ? Can I use the cuvettes more than once?

No. The electrical parameters provided by the Nucleofector™ Device are optimized for the cuvettes contained in the Nucleofector™ Kits. The cuvettes are single-use only. Using the cuvettes more than once will result in higher cell death and lower...

If I use Miltenyi paramagnetic beads for positive selection of cells, i.e. beads remain on the cell surface, does that influence Nucleofection results?

No, it seems that the beads bound to the cell surface do not have any influence on Nucleofection™. Larger magnetic beads (i.e. > 1µm) may disturb the Nucleofection™ process and negatively influence cell viability and efficiency.

What is the proof that DNA enters the nucleus during Nucleofection?

There are two facts that prove that the DNA is directly transported into the nucleus: 1.) Using the Nucleofector™ Technology, transfection of non-dividing cells is possible. With conventional non-viral transfection methods cell division is a...

How can I remove the cells from the cuvette after Nucleofector? Is there any alternative?

We recommend using the plastic pipettes provided in the kits. These pipettes prevent any damage to the cells and ensure a good cell recovery.

Do you have a protocol for the Nucleofection of Plasmodium berghei?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector™ Solution for your...

Is the age of the mice important for my mouse T cell Nucleofection?

Yes. We recommend using mice between 6-12 weeks. Using mouse T cells isolated from younger or older animals for Nucleofection™ may result in much lower transfection efficiencies and/or viabilities.
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