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I observed high mortality rates in mES cells after Nucleofection with a Cre recombinase expression vector. How can I increase the viability?

The High mortality after transfection might be due to the Cre recombinase itself, because the mammalian genome contains pseudo loxP sites (Thyagarajan B et al. (2000) Mammalian genomes contain active recombinase recognition sites. Gene....

I have a GFP construct that I use and usually get anywhere from 50-80% GFP positive cells using FACS analysis while I get only 30-40% using a GFP-NFAT fusion protein driven by the same promoter. Do you have any suggestions?

The Amaxa™ Nucleofection™ Conditions are optimised for each cell type, the solutions and programs are cell type dependent and not vector dependant. The expression of the fusion protein depends on the localization of the protein transcription,...

Can Lonza's antibiotics be used in clinical trials?

No. They are for research purposes only.

I currently work with some adherent cell lines, such as 293, which are grown in DMEM media. I have noticed that the mortality seems to be high after Nucleofection. Do you have any recommendations?

For some adherent cell lines we strongly recommend using RPMI instead of DMEM medium. As described in culture conditions of the cell line optimization kit protocol as well as the NIH3T3 (DSMZ) protocol, using RPMI instead of DMEM as "transfer and...

Can I use the Nucleofector Technology for RNAi applications? How do I start?

Yes. The Nucleofector™ Technology can be easily applied for any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the cell type specific Optimized Protocol for DNA vectors (cDNA, shRNA or miRNA expressing plasmids)...

Can I order any of the Nucleofector Kit components separately?

No. The Nucleofector™ Kits are only available as full kits.

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...

Who is the distributor for my country?

Please click on the link provided to search for your distributor.

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

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

Why do you have different Optimized Protocols for the Nucleofection of unstimulated and stimulated Human T cells?

We found that the stimulation of T cells not only changes the cells' function but also significantly alters the requirements for successful Nucleofection™. That's why Lonza developed two Optimized Protocols for transfection giving different...
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