Data Type


Category

+ Show All

Research Area

+ Show All
729 results sorted by

How can I remove the cells from the cuvette after Nucleofector? Is there any alternative?

We recommend using the plastic pipettes provided in the kits. These pipettes prevent any damage to the cells and ensure a good cell recovery.

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 cell...

What is the amount of DNA needed per well when using the 96-well Shuttle Device?

The amount depends on the cell type and on the DNA construct. As a rule of thumb 400 ng of DNA per well should be applied. However, for some cells or constructs even 100 ng may be sufficient.

What selection markers can I use for stable transfections?

The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.

What is the amount of siRNA needed per Nucleofection Reaction?

When performing siRNA-mediated knockdown experiments it is advisable to conduct a dose-response (concentration) analysis to determine the minimum siRNA concentration necessary for sufficient target knockdown on the mRNA, protein, or functional level....


Which pipette tips are compatible with the 96-well Shuttle Nucleocuvette Plates?

We recommend using special tips, e.g. epT.I.P.S™ (Eppendorf, Hamburg, Germany) or TallTips™ (Matrix Technologies, Hudson, NH, USA). You can contact our Scientific Support Team for a list of compatible tips, including tips for pipetting robots.

How often should I change the medium during selection of stable clones after Nucleofection?

Every 2-3 days. This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.

Can I freeze the 96-well Nucleocuvette Plates?

Yes. Freezing of the plates at -20°C does not alter their properties.

How many stable clones will I get from one transfection?

This depends on the respective cell type (transfection efficiency, viability and integration frequency). If it is important to you to know the frequency of stable integrants before carrying out your actual experiments, the frequency should be...
PAGE 8