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Batch to batch variation can be addressed by optimizing the transfection step in order to have a robust production process. Parameters that are critical for the transfection step are the transfection reagent used and whether the transfection reagent can form every time the right sized complexes to effectively deliver the DNA cargo into cells. The preparation of the transfection mix is therefore key to achieve the right sized complexes. Key parameters to consider are the amount of DNA per million of cells for transfection, the DNA to reagent ratio, the mixing strategy and length and the stability of the complexes. With Polyplus transfection reagents, these parameters have already been optimized to simplify your task. They can be further refined or adapted to your set up by contacting the Polyplus Scientific Support team. Lastly, it is important to make sure that the VCD at transfection remains the same from one batch to another.