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525 results sorted by
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We routinely use plasmids of 4-7 kb in our laboratories and plasmids up to approximately 20kb should not be a problem. Using plasmids larger than this will most likely result in lower transfection efficiency. Some preliminary results we have also...
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®
o
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s
?
The quality and the concentration of DNA used for Nucleofection® in general plays a central role for the efficiency of gene transfer. We strongly recommend using endotoxin free prepared DNA. Endotoxin free Kits are available from several...
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W
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y
?
The duration time of a program is dependent on the number of the program. The duration is also dependent on how long the Nucleofector® System has been turned on as well as the number of Nucleofections you have done in the last couple of...
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?
The recommended amount of DNA per 100 µl reaction is 0.5µg-5 µg. No carrier is needed.
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n
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n
c
e
?
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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r
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s
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s
?
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.
W
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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...
H
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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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K
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x
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s
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i
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n
?
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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t
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o
n
®
?
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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