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What is the proof that DNA enters the nucleus during Nucleofection?

There are two facts that prove that the DNA is directly transported into the nucleus: 1.) Using the Nucleofector™ Technology, transfection of non-dividing cells is possible. With conventional non-viral transfection methods cell division is a...

How can I remove the cells from the 100 uL Nucleocuvette Vessel after Nucleofection? Is there any alternative?

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

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 you...

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...

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 96-well...

Do you have any recommended methods for detaching human monocytes from the plate for assaying or before Nucleofection?

We have had good results by incubating the cells in ice cold PBS for 10 minutes and then rinsing the plates. Alternatively, the cells can be detached without a medium change by gently pipetting the cell suspension up and down. Our experience has also...

Can I use a Nucleofector I or Nucleofector 2b Device together with the 96-well Shuttle Device?

No, the 96-well Shuttle™ only works in conjunction with the Nucleofector™ IIs Device or our 4D-Nucleofector™ Device.

Is it possible to use different substrates (e.g. siRNA duplexes) with the 96-well Shuttle Device?

Yes. Transfection can be done with any substrate AND under exactly the same conditions. For example, the same conditions (96-well Shuttle™ Solution and program) are used for siRNA transfection or shRNA vectors. This greatly facilitates the...

What is the basic principle of the Nucleofector Technology?

The Nucleofector™ Technology is a novel transfection method especially designed for the needs of primary cells and difficult-to-transfect cell lines. It is a non-viral method based on a unique combination of electrical parameters and cell-type...
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