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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 method can I use to fix my HL-60 cells as shown in your optimized protocol?

The cells which are shown in the HL-60 optimized protocol are living cells and not fixed cells. However a good method for fixation of GFP is to use 3.5%-4% Paraformaldehyde in PBS. Please keep in mind that the fixation method is dependant on what...

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...

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.

Can I freeze the 96-well Nucleocuvette Plates?

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

Can I integrate the 96-well Shuttle Device into a liquid handling system?

Yes. The 96-well Shuttle™ Device is designed in such a way that it is addressable by robot grippers. Furthermore, the software contains the required interface to be connected with the LHS software. However, the practical implementation for each...

Can I use the Nucleofector 2b Solutions and Optimized Protocols with the 4D-Nucleofector X-Unit or the 96-well Shuttle Device?

No. The 4D-Nucleofector and 96-well Shuttle™ System uses specifically optimized electrical parameters and solutions. Five different 4D-Nucleofector or 96-well Shuttle™ Solutions are available for primary cells. For cell lines, three different...

Is your pmaxGFP control vector supplied with the Nucleofector Kits suitable for stable expression?

No. This vector is only supplied in our Nucleofector™ Kits as a positive control and cannot be used for selection because it does not contain a mammalian selection marker. The only resistance gene expressed by this vector is kanamycin which is only...
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