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The DNA bands on my agarose gel are sometimes wavy, but usually only in one or two lanes. What causes this?

Dried agarose on the comb teeth is a frequent cause of this problem. Prior to casting your gel, check the comb teeth for residual dried agarose.If not removed, this will attach to the newly cast agarose and fracture the well upon comb removal. This...

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.

Do you have any recommendation how to coat glass cover slips for neuron culture, e.g. for use with the Y unit or for post Nucleofection® culture in general?

Preparation of coverslips: At Lonza we used coverslips from Menzel (part of Thermo Scientific). We think that the coverslips have such a good quality that they don’t have to be cleaned prior to use.If you want to use other coverslips or prefer...

How much DNA should I load per well of an agarose gel?

The amount to load per well is variable. What is most important is how much DNA there is in the bands you wish to resolve. The least amount of DNA that can be consistently detected with ethidium bromide is about 10 ng. The most DNA you can have in a...

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.

At what voltage should I run an agarose gel?

We recommend running agarose gels at 4–10 volts/cm (cm is determined by measuring the interelectrode distance, not the gel length) under normal horizontal electrophoretic conditions.If the voltage is too high, band streaking, especially for DNA...

Do I have to apply the same program to each well of the 16-well Nucleocuvette®?

No, each well can be addressed separately. You may apply up to 16 different programs to one 16-well Nucleocuvette® Strip.

What is the difference between NuSieve 3:1 and NuSieve GTG Agaroses?

NuSieve™ 3:1 Agarose is a standard melting temperature agarose. The resolution range for NuSieve™ 3:1 Agarose is 50 bp–1000 bp. NuSieve™ 3:1 Agarose is designed for analytical electrophoresis; its high gel strength also makes it ideal for use in...

Can you use the AdipoRed Assay on cells other than preadipocytes?

Yes, the AdipoRed™ Assay Reagent can be used on many other cell types that accumulate triglycerides. It may have to be optimized for specific applications depending on the lipid that is stained.

How many 100 µl Nucleocuvettes® can be transfected in parallel using the 4D-Nucleofector® System?

The 4D-Nucleofector® X Unit can work with two 100µl Nucleocuvettes® in parallel. However, the operation software allows defining up to 50 Nucleocuvettes® per experiment. These cuvettes will be transfected by the system continuously in...
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