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W
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The generation of a stable clone often requires six months or more due to selection procedures and adaption to serum-free conditions after transfection.With the help of the Nucleofector® Technology suspension cells can be transfected directly in...
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This totally depends on the Nucleofector® system you are working with. While in the 100 µL reaction vessels usually 1 - 5 µg DNA is being used, for the 20 µL reaction vessels (e.g. X Unit, 96-well Unit and HT...
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According to our manufacturer, you can expect 12 million cortical neurons from an average E18-E19 rat pup.You can expect 4 million cortical neurons from an average mouse pup.
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The time course of plasmid expression after Nucleofection® might be different from the time course seen with other transfection methods. We recommend looking at different time points and start with the first time point as early as 4 to 6 hours post...
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In general the overall expression level differs between different cell types. Don't expect the same level of gene induction when working with primary cells. You might need to change to other DNA amounts or ratios.
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The original attenuation of the IRES sequence was performed to allow for a greater difference between the expression levels of the upstream gene of interest and the downstream reporter gene. If the downstream reporter gene, the product of less than...
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As the stability and half-life of various mRNAs and their protein products varies, it is important to empirically determine the best time points for assessing target knockdown. For example, it has been documented that in mammalian cells, mRNA...
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?
Yes. As the same protocol applies for any nucleic acid substrate (vectors or oligonucleotides) you can easily perform co-transfections either for transfection control or rescue experiments.
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We recommend using cuvettes with the 0.1 cm gap width.
W
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The reason is that insect cells are often incubated in very simple incubators that are not humidified (and without CO2 at 24°C to 28°C). Insect media typically do not need CO2 for buffering pH, so gas exchange is not required. To avoid evaporation of...
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