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What is the amount of siRNA needed per Nucleofection® Reaction?

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....

What protocol should I use for Nucleofection® of patient derived blood samples, e.g. leukemia or lymphoma cells?

Unfortunately, we do not have a ready-to-use protocol for Chronic lymphocytic leukemia (CLL) or other blood cell derived cancer cells. The cells often have a chromosomal aberration and show the properties of a cell line. Therefore we suggest to...

Are rat and mouse hepatocytes transfected by Nucleofection® still functional?

Yes, transfected mouse hepatocytes show albumin synthesis rates comparable to those of untransfected cells. Moreover the morphology and polarization of rat hepatocytes is not affected by Nucleofection®.

Which pipette tips are compatible with the 96-well Shuttle® Nucleocuvette® Plates?

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.

Can I freeze the 96-well Nucleocuvette® Plates?

Yes. Freezing of the plates at -20°C does not alter their properties.

Can I integrate the 96-well Shuttle® Device into a liquid handling system?

Yes. The 96-well Shuttle® Device is designed in such a way that it is addressable by robot grippers. Furthermore, the software contains the required interface to be connected with the LHS software. However, the practical implementation for each...

Can I use the Nucleofector® 2b Solutions and Optimized Protocols with the 4D-Nucleofector® X-Unit or the 96-well Shuttle® Device?

No. The 4D-Nucleofector and 96-well Shuttle® System uses specifically optimized electrical parameters and solutions. Five different 4D-Nucleofector or 96-well Shuttle® Solutions are available for primary cells. For cell lines, three...

Generation of a stable cell line: A positive control plasmid works, but I cannot obtain stable clones with my construct of interest. What could be the reason?

One reason could be that your gene product is toxic to the cells. We recommend testing the construct for expression of the gene of interest by transient Nucleofection® of your cells. If the proportion of expressing cells drops between 4 and 48 hours...

How long does it take to obtain stable transfectants?

Depending on individual cell type and doubling rate, selection of stable transfectants will take between 7 and 28 days. Expansion and characterization of single cell clones will take several weeks in addition.

Can I use a Nucleofector® I or Nucleofector® 2b Device together with the 96-well Shuttle® Device?

No, the 96-well Shuttle® only works in conjunction with the Nucleofector® IIs Device or our 4D-Nucleofector® Device.
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