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Are your Nucleofector Solutions checked for RNase activity during your QC process?

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...

What kind of buffer do you recommend for resuspending my siRNA?

Please use the buffer that is recommended by your siRNA manufacturer.

What stock concentration of Propidium Iodide does Lonza use for FACS analysis?

We use a stock concentration of 100 µg/ml in PBS. This is diluted 1:300 – 1:400 into the sample.

What kind of protocol you recommend for stimulation of human T cells?

For getting the results we present in our datasheet, we recommend the follwing strategy: Cells should be stimulated and cultivated for 3 days before Nucleofection, preferentially on plates coated with 1 µg/ml anti-CD3 and 2 µg/ml anti-CD28 antibody....

Are there any effects on the differentiation of neural stem cells following Nucleofection?

The ability of transfected rat or mouse neural stem cells to be subsequently differentiated into a variety of cell types is well documented in: Olig2 Overexpression Induces the In Vitro Differentiation of Neural Stem Cells into Mature...

Can calcium influx issues affect viability of human Mesenchymal Stem Cells (hMSC)?

Yes. When cells are transfected using Nucleofection™, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection™ in media that...

Can calcium influx issues affect viability or differentiation in Stem Cells after Nucleofection?

When cells are transfected by Nucleofection™, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection™ in medias that contain high...

What are the requirements for direct Nucleofection of mRNA for protein expression?

The mRNA should be capped and polyadenylated. The conditions of Nucleofection™ will be the same as for DNA with the particular cell type, i.e. follow the same protocol and use the same program, except one will likely need to add a much higher amount...

Do you have any siRNA Nucleofection results using concentrations lower than 50nM?

Yes. Please click on the attached Technical Reference Guide: "Designing an RNAi Experiment Using Nucleofection™". On page 3 you can find a table with some examples.

The animal facility which isolates my hepatocytes is at a different location, should I be concerned about transport time?

Isolated hepatocytes can be transported in Krebs-Henseleit Buffer or supplemented William's E Medium. Long transport time for isolated hepatocytes, (in excess of one hour) can cause a decrease in efficiency of Nucleofection™, but does not seem to...
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