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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...
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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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DNA amount and quality are very critical for Nucleofection™ of Dendritic Cells (recommended 0.5-1µg; maximal 2 µg). LPS (lipopolysaccharides) have a strong negative effect on cell viability. Please make sure that all components you use for dendritic...
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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...
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Yes. We do test the Solutions with a real time RT-PCR based assay. This assay basically measures the quantity of an RNA template after incubation in our solution. In case there is RNAse contamination the template would be digested which would be...
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The pmaxGFP control DNA is not suitable for making stable clones because it lacks a selectable marker for mammalian cells. The kanamycin resistance expressed by this plasmid is only suitable for bacterial selection.
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The expiration dates are located on the outside label of the kit box and on the label of the solution box.
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Please use the buffer that is recommended by your siRNA manufacturer.
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We use a stock concentration of 100 µg/ml in PBS. This is diluted 1:300 – 1:400 into the sample.
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As long as the supplement was not added to the Nucleofector™ Solution, then there is no risk of any damage to the solutions. Even long-term storage of several months did not alter the performance of the Nucleofector™ Solution. However, if the...
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