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M
y
N
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As long as the supplement was not added to the Nucleofector™ Solution, then there is no risk of any damage to the solutions. Even long-term storage of several months did not alter the performance of the Nucleofector™ Solution. However, if the...
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In order to avoid cell compaction in the pellet and difficulties in cell resuspension, we recommend using round bottom 2 ml vials or 50 ml BD Falcon™ tubes for all centrifugation steps prior to Nucleofection™.
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The Amaxa™ Nucleofection™ Conditions are optimised for each cell type, the solutions and programs are cell type dependent and not vector dependant. The expression of the fusion protein depends on the localization of the protein transcription,...
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For some adherent cell lines we strongly recommend using RPMI instead of DMEM medium. As described in culture conditions of the cell line optimization kit protocol as well as the NIH3T3 (DSMZ) protocol, using RPMI instead of DMEM as "transfer and...
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N
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c
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f
e
c
t
i
o
n
™
?
Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).
C
a
n
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?
H
o
w
d
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s
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t
?
Yes. The Nucleofector™ Technology can be easily applied for any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the cell type specific Optimized Protocol for DNA vectors (cDNA, shRNA or miRNA expressing plasmids)...
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?
The RosetteSep Kit is less expensive than the MACS Selection Kit. The MACS Selection Kit results in a higher purification (up to 97-98%) of the specific cell population.
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w
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h
t
h
e
N
u
c
l
e
o
f
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™
?
We recommend allowing the transfected hepatocytes to recover in Plating medium for 72 hours post-Nucleofection™. For induction studies (e.g. CYP3A4), the Plating medium is replaced with maintenance medium (William's E without FCS and EGF) and...
C
a
n
I
o
r
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e
r
a
n
y
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f
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N
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f
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™
K
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t
c
o
m
p
o
n
e
n
t
s
s
e
p
a
r
a
t
e
l
y
?
No. The Nucleofector™ Kits are only available as full kits.
D
o
y
o
u
r
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c
o
m
m
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n
d
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s
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n
b
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f
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r
e
N
u
c
l
e
o
f
e
c
t
i
o
n
™
?
For obtaining highly purified cell populations, we always recommend using the MACS™ Selection Kit (Miltenyi Biotec GMbH).
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