Tricks in choosing diluent for GenJet™, LipoD293™ & PolyJet™ DNA transfection reagents.

Tricks in choosing diluent for GenJet™, LipoD293™ & PolyJet™ DNA transfection reagents.

For efficient formation of transfection complex, to choose an appropriate diluent which is used for diluting DNA transfection reagent and DNA is essential. In addition to temperature, incubation time, the nature of diluent is very important for complete formation of transfection complex, thus significantly affecting DNA transfection efficiency. Per our validation data, an appropriate diluent can be to 50 times more efficient than a "wrong" diluent. What's more important is researchers often neglect the importance of diluent and fail to recognize role of diluent in the process of transfection complex formation.

Follow the following technical tips to choose the proper diluent for GenJet™, PolyJet™ and LipoD293™ DNA transfection reagents:
1. For complete formation of transfection complex, the diluent must be serum-free and protein free.
2. Try to use a diluent which is closest to the composition of cell culture medium. For instance, if you are using DMEM (supplemented with serum and antibiotics) to grow HEK293 cells, the best diluent is serum-free and antibiotics-free DMEM. If you are using RPMI-1640 Medium (supplemented with serum and antibiotics) to grow Hela cells, the best diluent is serum/antibiotic-free RPMI-1640 medium.
3. Never use a medium which contains unknown components. For example, never use Opti-MEM™ to dilute GenJet™, PolyJet™ and LipoD293™ DNA transfection reagents. Per our validation data, diluting our transfection reagents and DNA with Opti-MEM™ at least sacrificed 50% transfection efficiency on HEK293, Hela, CHO and NIH 3T3 cells. Also Opti-MEM™ may contains low level of serum which will greatly compromise transfection efficiency.
4. Basically serum-free DMEM with high glucose is pretty good choice. It is very easy accessible and compatible with other cell growth mediums. If you have trouble with lower transfection efficiency, you may think about changing diluent to serum-free DMEM with high glucose. If you do not know how to choose a proper diluent, you can try serum-free DMEM with high glucose first which usually gives you very good transfection efficiency.


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