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526 results sorted by
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The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.
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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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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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Antibiotics for selection should be added 24-48 h after Nucleofection®.
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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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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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The culture medium should be changed every 2-3 days during the selection process.This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.
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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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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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This depends on the respective cell type (transfection efficiency, viability and integration frequency). If it is important to you to know the frequency of stable integrants before carrying out your actual experiments, the frequency should be...
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