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The mRNA should be capped and polyadenylated. The conditions of Nucleofection® will be the same as for DNA with the particular cell type, i.e. follow the same protocol and use the same program, except one will likely need to add a much higher...
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Nucleofector I/II/2bWe have excellent data for the Nucleofection™ of Plasmodium berghei from the group Chris J. Janse and Andrew P. Waters from the University of Leiden. Their protocol describes a transfection efficiency of 10-3 – 10-4. With this...
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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...
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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...
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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...
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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 Primary Cell...
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No, the programs C-20 and C-020 are identical regarding their electrical parameters. The letter minus a two-digit number is the nomenclature of the Nucleofector™ I. Since we have launched the Nucleofector™ II we changed it to a letter minus a three...
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For Nucleofectio® 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...
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The results obtained using 7AAD staining are much clearer than those obtained using propidium iodide (PI). Using flow cytrometry stained and unstained, transfected and non-transfected populations can be separated much more easily.
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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™...
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