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DNA-purity and Nucleofection®: Can low A260:A280 ratios lead to both reduced transfection efficiency and cell viability?

Yes. To check the quality of your DNA we strongly recommend determining the A260:A280 ratio. It should be ideally 1.8 - 2.0 for a good DNA preparation, but least 1.6.

Is it possible to quantify the mycoplasma with the MycoAlert® Mycoplasma Detection Assay?

No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.

What is the advantage of using conductive polymer as electrode material for the 4D-Nucleofector® X-Unit and 96-well Unit?

With conductive polymer electrodes the release of metal ions into the cell suspension is avoided. Conductive polymer electrode Nucloecuvette® Vessels are used in the 4D-Nucleofector® X-Unit and 96-well Unit?

Can nicked DNA lead to reduced transfection efficiency in Nucleofection® experiments?

Yes. You should verify the integrity of your DNA on an agarose gel to see if it is degraded. Compare undigested plasmid to plasmid DNA digested with a suitable single cut restriction enzyme to linearize. Supercoiled plasmid will run faster than...

When performing the MycoAlert® Mycoplasma Detection Assay, why is it necessary to spin out the cells?

It is highly recommended that the samples are spun down prior to testing. If cells are present in the assay they will contribute to the initial reading producing a higher background. This in turn can decrease the difference between the background and...

Can I use the settings recommended for the Nucleofector® I/II or 2b System with my 4D-Nucleofector® System?

No, the 4D-Nucleofector® is optimized for conductive polymer electrodes, while the Nucleofector® I/II or 2b System is working with electrodes made of aluminium.We recommend doing a quick cell line optimization using our Cell Line...

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...

Can I reuse the 100µl Nucleocuvette® Vessel?

We do not recommend reusing the conductive polymer cuvettes as performance decreases when cuvettes are used more than once.

How long is the MycoAlert® Mycoplasma Detection Assay signal stable?

Please follow the protocol exactly as written – add reagent, wait 5 min and read, add substrate, wait 10 minutes and read again. 

How many cells do I have to use for each well of the 16-well Nucleocuvette® Strip?

This depends on the cell type you are working with. In our Optimized Protocols the cell numbers range from 2.5 x 10^4 cells (mouse DC) up to 1x 10^6 cells (human T cells). Detailed information is provided in our 4D-Nucleofector® Optimized Protocols.
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