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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.

Why is the Nucleofector® Technology ideal for protein production from stable clones ?

The generation of a stable clone often requires six months or more due to selection procedures and adaption to serum-free conditions after transfection.With the help of the Nucleofector® Technology suspension cells can be transfected directly in...

Do you have any information regarding differentiation of mouse ES cells after Nucleofection® with large plasmids?

It is unlikely that plasmid size would affect in vitro differentiation capability. Gene targeting constructs are often as large as 20 kb and if treated well the ES cells are perfectly capable of generating germ line chimaeras afterwards (indicating...

What kind of cuvettes can one use for the transformation of bacteria using the Nucleofector® I/II/2b?

We recommend using cuvettes with the 0.1 cm gap width.

Why does Lonza recommend using non-ventilated caps when culturing insect cells?

The reason is that insect cells are often incubated in very simple incubators that are not humidified (and without CO2 at 24°C to 28°C). Insect media typically do not need CO2 for buffering pH, so gas exchange is not required. To avoid evaporation of...

For Lonza's animal neurons, how many cortical neurons can I expect from a single pup?

According to our manufacturer, you can expect 12 million cortical neurons from an average E18-E19 rat pup.You can expect 4 million cortical neurons from an average mouse pup.

Does the Mouse T cell protocol work for stimulated mouse T cells?

Yes. Stimulate the cells with anti CD3 and anti CD28 antibody for 24 hours. Use the optimized protocol. Often times, this will result in higher transfection efficiency (60-70% is possible). Alternatively, you can stimulate with ConA and IL-2 for 2-3...

What is the ratio of Supplement to Nucleofector® Solution?

The ratio is 1:4,5 (500 µL of supplement is added to 2.25 mL of Nucleofector® Solution).For a single 100 µL reaction, use 18 µL of supplement plus 82 µL of solution to make the 100 µL.

Can I apply more than one program (transfection condition) per plate with the 96-well Shuttle® System?

With the 96-well Shuttle® system, it is possible to apply up to 96 different programs (transfection conditions) per 96-well Nucleocuvette® Plate. Therefore, optimization of Nucleofection® Conditions for a particular cell line can be...

What cell number do I need per well when using the 96-well Shuttle® Device or the 4D-Nucleocuvette® Stripes?

The cell number is very much dependent on the cell type you use. For high throughput Nucleofection®, we usually start with one fifth of the optimal amount recommended for the standard Nucleofection® in the 100µl cuvette. Currently the absolute...
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