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In my Nucleofection Experiment I see lower expression with my IRES construct in comparison with you pmax-GFP control. Do you have any information about the different expression profiles?

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...

Do we sell the Nucleofector Solution (or any other component of the Nucleofector Kit) separately?

We don't sell the individual components of the kits separately because our product line is a kit concept line to ensure the use of Amaxa™ certified products with quality controlled lot numbers that guarantee optimal results.

What are the key things in a Nucleofection that one can do to maximize viability?

Pay careful attention to centrifugation speeds prior to Nucleofection™; be sure not to centrifuge the cells at higher than 90xg. Any trypsinization should use the minimal amount of reagent and time necessary in order to minimize stress to the cells,...

Do your Clonetics Rat and Mouse Astrocytes require an extracellular matrix for plating?

No, they simply need to be seeded onto tissue culture treated plastic. For best performance, we recommend to use positively charged plasticware

What are the best time points for analysis of my siRNA experiments?

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...

What age are the rats that your Clonetics Rat DRGs are isolated from?

They R-DRG-505 are isolated from neonatal, about 2-3 days old. The R-eDRG-515 are isolated from embryonic rats (E14-15).

Does Lonza have any recommendations for siRNA controls to try in human T cells?

The following controls have been found to work well in the Nucleofection™ of human T cells: GAPDH, Vimentin, Cycophilin B, and Beta-Actin. We do not recommend the following for use as controls in human T cells: PLK and Laminin A/C.

Can I co-transfect oligonucleotides and plasmids with the Nucleofector Technology?

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.