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DNA HTS: jetPEI®

  • Fast and efficient methods to transfect cells for HTS
  • Exceptionally reproducible results
  • Well-suited for automated approaches
  • Compatible with serum and antibiotics
  • Reverse, batch & forward protocols available

jetPEI® transfection reagent is a linear polyethylenimine derivative, free of components of animal origin, providing highly effective and reproducible gene delivery.
jetPEI® transfection reagent is therefore particularly well suited for automated or manual HTS (High Throughput Screening) with three protocols available: reverse, batch and forward.

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.

More information:

Expand All Tabs
Catalog Number Amount of reagent Amount of NaCl solution
101-01N 0.1 mL 5 mL
101-10N 1 mL 50 mL
101-10 1 mL -
101-40N 4 x 1 mL 4 x 50 mL
101-40 4 x 1 mL -
101B-010N 10 mL 2 x 250 mL
101B-010 10 mL -

NaCl complex-formation solution included (adapted to proper complex-formation, as indicated in the protocol).

One ml of jetPEI® is sufficient to perform 2 000 transfections in 96-well plates.

Bulk quantities are available upon request (50 mL, 100 mL and 1L).

Please contact us.

 

The reverse protocol is the most appropriate when transfecting a pool of genes, such as a DNA library (Fig. 1). In this protocol, the jetPEI®/DNA complexes are prepared or deposited in the wells prior to addition of the cells. Complexes are stable for up to 4 hours (Fig. 2).

The batch protocol was developed to prepare a homogeneous pool of transfected cells. For this purpose, the cells are transfected just after trypsinization, while still in suspension. This protocol is prefered for drug screening applications and allows rapid processing, one day faster than the forward protocol.

In the forward protocol, the cells are split the day before transfection and the jetPEI®/DNA complexes are added to the adherent cells.

HTS-jetPEI-Fig1

 

 

Figure 1. jetPEI® reverse transfection protocol for HTS application.

 

Complexes formed with the water-soluble polymer jetPEI® and DNA allow efficient transfection for up to 4 hours, in contrast to lipid-based reagents and calcium phosphate. Thus they allow plenty of time to dispense the complexes into the plates (Fig. 2).

HTS-jetPEI-Fig2 Figure 2. Effect of complex formation incubation time on transfection efficiency with jetPEI®. HEK 293 cells were transfected in 96-well plates with pCMVLuc and jetPEI™ following the reverse transfection protocol. Luciferase activity was measured after 24 h.

 

HTS DNA transfection using jetPEI® gives highly consistent transfection efficiency from batch-to-batch (Fig. 3).

HTS-jetPEI-Fig3 Figure 3. Batch-to-batch reproducibility using jetPEI®. For each lot, HeLa cells were transfected in triplicate in the presence of serum in triplicate using the standard protocol in 24-well plate.

 

jetPEI® successfully delivers genes to various adherent and non-adherent cell lines, as well as primary cells (Table 1). Over 550 publications using jetPEI® can be found in the Product Citation Database on the Polyplus website. In addition, a Cell Transfection Database gives specific transfection conditions for over 400 cell lines and primary cells.

HTS-jetPEI-Table1 Table 1. Some common cell lines and primary cells successfully transfected using jetPEI®.

 

jetPEI® was compared to several other popular transfection reagents (Fig. 4). jetPEI® was found to offer the best performance: high efficiency and low variability (small standard deviation).

HTS-jetPEI-fig4 Figure 4. Transfection efficiency of a series of commercial reagents. HeLa cells were transfected in 24-well plates in the presence of 10% serum, using 1 µg pCMV-luciferase according to the manufacturers’ protocols. Luciferase expression was measured 24 h after transfection.