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Yes, DsRed is much less bright than GFP and is more difficult to detect by FACS and microscopy because it bleaches quickly.
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The pmaxGFP™ Vector provided in our Nucleofector™ Kits is not expressed in insect cells. We strongly recommend an insect expression vector encoding a fluorescent protein or lacZ reporter as a positive control for your experiments [e.g. Novagen™’s...
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It is unlikely that plasmid size would affect in vitro differentiation capability. Gene targeting constructs are often as large as 20 kb and if treated well the ES cells are perfectly capable of generating germ line chimaeras afterwards (indicating...
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Yes. Stimulate the cells with anti CD3 and anti CD28 antibody for 24 hours. Use the optimized protocol. Often times, this will result in higher transfection efficiency (60-70% is possible). Alternatively, you can stimulate with ConA and IL-2 for 2-3...
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With the 96-well Shuttle™ Device, it is possible to apply up to 96 different programs (transfection conditions) per 96-well Nucleocuvette™ Plate. Therefore, optimization of Nucleofection™ Conditions for a particular cell line can be easily performed...
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The cell number is very much dependent on the cell type you use. For high throughput Nucleofection™, we usually start with one fifth of the optimal amount recommended for the standard Nucleofection™ in the 100µl cuvette. Currently the absolute cell...
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The amount depends on the cell type and on the DNA construct. As a rule of thumb 400 ng of DNA per well should be applied. However, for some cells or constructs even 100 ng may be sufficient.
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R
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When performing siRNA-mediated knockdown experiments it is advisable to conduct a dose-response (concentration) analysis to determine the minimum siRNA concentration necessary for sufficient target knockdown on the mRNA, protein, or functional level....
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P
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We recommend using special tips, e.g. epT.I.P.S™ (Eppendorf, Hamburg, Germany) or TallTips™ (Matrix Technologies, Hudson, NH, USA). You can contact our Scientific Support Team for a list of compatible tips, including tips for pipetting robots.
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Yes. Freezing of the plates at -20°C does not alter their properties.
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