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Transient transfections were performed using 40 kDa linear PEI (Polysciences) as transfection reagent. 2 μg DNA was combined with 100 μl Sf-900 II SFM medium and the same was done for 10 μg PEI. DNA/PEI complex formation was performed by combining both mixes, followed by incubation for 15 min at room temperature. |
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| Protocol tips |
| Transient transfections were performed using 40 kDa linear PEI (Polysciences) as transfection reagent. 2 μg DNA was combined with 100 μl Sf-900 II SFM medium and the same was done for 10 μg PEI. DNA/PEI complex formation was performed by combining both mixes, followed by incubation for 15 min at room temperature. |
Publication protocol
Sf9 insect cells (Novagen/Merck Millipore) were amplified and transferred to 6-well plates in Sf-900 II SFM medium (Thermo Fisher Scientific) supplemented with 5% fetal bovine serum. Transient transfections were performed using 40 kDa linear PEI (Polysciences) as transfection reagent. 2 μg DNA was combined with 100 μl Sf-900 II SFM medium and the same was done for 10 μg PEI. DNA/PEI complex formation was performed by combining both mixes, followed by incubation for 15 min at room temperature. After replacing the cell medium with Sf-900 II SFM, the transfection mix was added drop-wise to the cells. Harvesting was performed three days after transfection.
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