Technical tips for optimizing PolyJet reagent.

PolyJet DNA Transfection Reagent is highly efficient for transfecting epithelial and RAW 267.4 cells without any significant toxicity. The following are some technical points for maximizing efficiency of the PolyJet transfection reagent.
1. DNA amount and quality: DNA purity is essential for high efficiency. DNA prepared from E Coli. must be 1.80 at A260/280 or higher. For a 6-well plate, ~1.0 µg of DNA per well is usually sufficient. For other cell culture formats, adjust the amount of DNA with consideration to the surface area of the cell culture dish.
2. Ratio of PolyJet/DNA: For epithelial and RAW 267.4 cells, we found a ratio of 3:1 had excellent results.
3. Diluent: The diluent used to dilute DNA and PolyJet reagent must be serum free, such as serum-free DMEM with high glucose. Opti-MEM is NOT a good choice because it may affect formation of the transfection complex, so it is not recommended for this step.
4. Transfection with serum: We performed transfection with and without serum/antibiotics and failed to find a significant difference in transfection efficiency. PolyJet reagent is therefore NOT interfered by serum/antibiotics.

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