DS- Fast and efficient methods to transfect cells for HTS
- Compatible with serum and antibiotics
- Robust jetPEI/DNA complexes
- Exceptionally reproducible results
- Well-suited for automated approaches
jetPEI™ transfection reagent is particularly well suited for automated or manual HTS (High Throughput Screening) as transfection with jetPEI™ is exceptionally efficient and reproducible. Three HTS protocols are available depending on the experiment: forward, batch or reverse.
INTELLECTUAL PROPERTY
The use of polyethylenimine (PEI) or polypropylenimine (PPI) or cationic polymers similar in structure thereto for transfecting cells, as well as compositions comprising these cationic polymers and at least one nucleic acid, are the subject matter of U.S. Patent No. 6,013,240, EP Patent No. 0770140 and foreign equivalents, for which Polyplus-transfection™ is the worldwide exclusive licensee.
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Ordering informationjetPEI™ transfection reagent can be used efficiently for automated or manual HTS (High Throughput Screening) with three different protocols depending on the experiment:
- Forward HTS protocol
- Batch HTS protocol
- Reverse HTS protocol
In the forward protocol, the cells are split the day before transfection and the jetPEI™DNA complexes are added directly to adherent cells. Since the complexes are stable and the complexation time fl exible, the forward protocol is the most appropriate for routine HTS application.
The batch protocol was developed to prepare a homogenous pool of transfected cells. For this purpose, the cells are transfected straight after trypsinization while still in suspension. This protocol is best suited for drug screening applications and allows a rapid processing by saving one day compared to the forward protocol.
When transfecting a pool of genes, such as a DNA library, the reverse protocol is the most appropriate. In this protocol, the jetPEI™DNA complexes are prepared or deposited in the wells prior to adding the cells.This reverse protocol is the most commonly used in HTS.
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Figure 1. Forward, batch and reverse transfection protocols available using jetPEI™for HTS application
In contrast to lipid transfection reagents and calcium phosphate, the water-soluble polymer jetPEI™ forms stable complexes with DNA allowing ample time for time consuming automated approaches (Fig. 2).
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Figure 2. Effect of incubation time for complex formation on transfection efficiency. HeLa cells were transfected with pCMVLuc using jetPEI™. Luciferase activity was measured after 24 h. |
The incubation time for jetPEI™/DNA complex formation from 10 min up to 2 hours does not affect transfection efficiency, allowing plenty of time to dispense the complexes into the plates.
As a result of the stability of the DNA/jetPEI™ complexes, the protocol is very robust, showing no variation in transfection efficiency between the first and the last well of a series (Fig. 3) and from one experiment to the next.
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Figure 3. ß-galactosidase assay after transfection using the HTS forward protocol in 96-well plates. BHK-21 cells were transfected with pCMVLacZ using jetPEI™. Identical gene expression levels were observed for all wells. |
When compared to other commercial reagents, jetPEI™ was found the most efficient, showing the lowest variability in transfection efficiency (Fig. 4).
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Figure 4. Comparison of jetPEI™ efficiency with other commercial reagents. HeLa cells were transfected withpCMV-Luc according to the manufacturers’ protocols. Luciferase expression was measured after 24 h. |
