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Ingenio® EZporator® Electroporation System
Product User Manual
| Instructions for MIR 51000
EZporator® System Protocols (Continued)
Instructions for Use with Ingenio® Electroporation Solution and Kits
The Ingenio® Electroporation Solution and Kits provide a universal, high efficiency,
low toxicity solution for electroporating nucleic acids (i.e. DNA, RNA, oligonucleo-
tides, etc.) or other molecules into mammalian and insect cell types.
Transient plasmid DNA or siRNA electroporation protocol
A. Plate cells
1. Approximately 18
-
24 hours before electroporation, passage cells to attain an
optimal cell density at the time of electroporation (i.e. 70
-
90% confluent for
most cell types).
For adherent cells
: Plate cells at a density of 0.8
-
3.0 × 10
5
cells/ml
For suspension cells
: Seed cells at a density of 1
-
2 × 10
6
cells/ml
2. Incubate cell cultures overnight.
B. Prepare Ingenio® Solution/nucleic acid/cell mixture for electroporation
(immediately before electroporation)
1. Warm Ingenio® Electroporation Solution, trypsin
-
EDTA (if applicable) and
complete growth medium to room temperature.
2. Harvest and count cells to determine cell density (cells/ml).
3. Determine the total electroporation volume required to perform the desired
number of electroporations.
For 0.2 cm cuvettes:
Multiply number of electroporations by 0.1 ml
For 0.4 cm cuvettes:
Multiply number of electroporations by 0.25 ml
4. Calculate the cell volume required for all electroporations:
For adherent cells
: Use a final cell density of 1
-
5 × 10
6
cells/ml
For suspension cells
: Use a final cell density of 10 × 10
6
cells/ml
5. Pipette the cell volume (from step B4) of harvested cells into a new tube and
centrifuge at 300
× g
for 5 minutes. Aspirate the supernatant.
6. During centrifugation, add pre
-
warmed complete culture medium to a new
culture dish to accept cells following electroporation.
7. After centrifugation, resuspend cells in Ingenio® Electroporation Solution us-
ing the volume determined in step B3.
(Cont. on next page)
Mirus Bio
LLC
U.S. Toll Free: 844.MIRUSBIO
(844.647.8724)│
Direct: +1.608.441.2852
Final Cell Density (cells/ml) × Volume from Step B3 (ml)
Harvested Cell Density from Step B2 (cells/ml)
Cell Volume (ml) =
