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Ingenio® EZporator® Electroporation System
Product User Manual
| Instructions for MIR 51000
Optimization
Electroporation parameters (e.g. cell density, nucleic acid amount, voltage) should
be optimized for each cell type to ensure best possible transfection efficiency and
cell viability. The suggestions below yield high efficiency electroporation of most
mammalian and insect cell types.
Cell Density and Passage Number
•
Cell division.
Passage cultures approximately 18
-
24 hours before electro-
poration to ensure that cells are actively dividing and reach the appropriate
density (i.e. 70
-
90% confluent for adherent cells or 2
-
4 × 10
6
cells/ml for sus-
pension cells) at the time of harvesting for electroporation. In general, cells
should be in mid
-
logarithmic growth for optimal electroporation.
•
Cell density at electroporation.
Determine the optimal cell density for each cell
type to maximize electroporation efficiency. The optimal cell density for elec-
troporation is typically between 1
-
10 × 10
6
cells/ml. Higher cell densities are
typically recommended for suspension cells (e.g. 1 x 10
7
cells/ml), whereas the
recommended range for adherent cells is 1
-
5 × 10
6
cells/ml.
•
Cell passage number.
Use of very low or very high passage cells may affect
experimental results. Use cells of similar passage number for experimental
reproducibility.
Nucleic Acid Purity and Concentration
•
Plasmid DNA.
Use highly purified, sterile and contaminant
-
free DNA for elec-
troporation. Plasmid DNA preparations that are endotoxin
-
free and have
A260/280 absorbance ratio of 1.8
-
2.0 are desirable. DO NOT use DNA prepared
using miniprep kits or DNA that is less than 0.5 mg/ml in concentration. To
determine the best plasmid DNA concentration for electroporation, try DNA
concentrations in the range of 5
-
50
μ
g/ml of final electroporation volume.
•
siRNA.
Use siRNA that is highly pure, sterile and the correct sequence. Try siR-
NA in the range of 250
-
750 nM (final concentration in cuvette) to determine
the best siRNA concentration for electroporation.
Mirus Bio
LLC
U.S. Toll Free: 844.MIRUSBIO
(844.647.8724)│
Direct: +1.608.441.2852
