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What is the basic principle of the Nucleofector Technology?

The Nucleofector™ Technology is a novel transfection method especially designed for the needs of primary cells and difficult-to-transfect cell lines. It is a non-viral method based on a unique combination of electrical parameters and cell-type...

Why is the Nucleofector Technology ideal for primary cells and difficult-to-transfect cell lines?

There are several reasons to choose the Nucleofector™ Technology for your gene transfer experiments. One is the direct transfer of DNA to the cell nucleus. This makes gene expression independent from cell division. Therefore, the technique is the...

What optimization is necessary to get the Nucleofector Technology to work?

None. For each cell type in our product list we offer a cell-type specific solution and thoroughly optimized electrical parameters. These are already pre-programmed in the Nucleofector™ Device. Additionally our optimized protocols give you...

What can I do when there is no optimized protocol available for my cell of interest?

"Lonza is constantly developing new optimized kits and protocols for an increasing number of primary cells and cell lines. In order to obtain the latest information, check our website or contact our Scientific Support Team.We are offering Cell Line...

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...

What is the best way to establish the Nucleofector Technology in my lab?

We strongly recommend establishing the Nucleofector™ Technology with the positive control vector pmaxGFP™ provided in our kits. pmaxGFP encodes the green fluorescent protein (GFP) from Copepod Pontellina sp. Just like eGFP expressing cells, maxGFP™...

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.

Is there a special recommendation on Human T cell enrichment before Nucleofection?

For Nucleofection™ it is preferable to use Human T cell populations enriched by magnetic separation. For example, MACS™ Microbeads (Miltenyi) or by a rosetting method. For magnetic separation we recommend negative selection or detaching beads after...

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™...

What method can I use to fix my HL-60 cells as shown in your optimized protocol?

The cells which are shown in the HL-60 optimized protocol are living cells and not fixed cells. However a good method for fixation of GFP is to use 3.5%-4% Paraformaldehyde in PBS. Please keep in mind that the fixation method is dependant on what you...
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