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524 results sorted by
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C
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No. The electrical parameters provided by the Nucleofector® System are optimized for the cuvettes contained in the Nucleofector® Kits. The cuvettes are single-use only. Using the cuvettes more than once will result in higher cell death and lower...
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?
No, it seems that the beads bound to the cell surface do not have any influence on Nucleofection®. Larger magnetic beads (i.e. > 1µm) may disturb the Nucleofection® process and negatively influence cell viability and efficiency.
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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...
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?
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.
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?
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...
D
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N
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®
?
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...
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q
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?
For the preparation of mouse spleen cells we recommend cutting the spleen once and passing the tissue through a 100µM cell strainer or steel mesh using a plunger. The cells are flushed into a petri dish containing PBS. In order to remove fat, cell...
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®
?
We only recommend the depletion method. For Example, the RosetteSep™ Isolation Kit for human monocytes [Stem Cell Technologies, Cat. No 15028]. The advantage of depletion is that the monocytes are left untouched by antibodies during the process.
D
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?
The PBS should be calcium and magnesium free to prevent clumping of cells. We routinely use PBS without calcium and magnesium for example Lonza Cat. No. BEBP17-516Q
I
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T
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N
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f
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c
t
i
o
n
®
?
Yes. We recommend using mice between 6-12 weeks. Using mouse T cells isolated from younger or older animals for Nucleofection® may result in much lower transfection efficiencies and/or viabilities.
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