选择何种稀释液稀释DNA及转染试剂，对于制备有效的转染复合物至关重要。除了温度，孵育时间外，稀释液的性质对于制备高效的转染复合物亦非常重要，同样影响DNA转染效率。根据我们的实验数据，使用合适的稀释液得到的转染效率是使用错误稀释液转染效率的至少50倍。更为重要的是，实验者总是忽视稀释液的重要性，甚至一直未曾意识到正确的稀释液对形成有效转染复合物的重要性。
如下实验技巧可以帮助您为GenJetTM，LipoD293TM&PolyJetTMDNA转染试剂选择合适的稀释液。
1、为了制备更有效的转染复合物，稀释液中千万不要含有血清、蛋白。
2、尽量使用接近细胞培养基成分的稀释液，例如：若是你使用DMEM（supplemented with serum and antibiotics）培养HEK293细胞，那么，最合适的稀释液是不含血清、抗生素的DMEM：若是您使用RPM1-1640（supplemented with serum and antibiotics）培养Hela细胞，那么最合适的稀释液是不含血清、抗生素的RPM1-1640。
3、请勿使用含有未知成分的培养基。例如，切勿使用Opti-MEMTM稀释GenJet™、PolyJet™及LipoD293™ DNA转染试剂。根据我们的实验数据，若是使用Opti-MEM稀释转染试剂及DNA，那么用于HEK293，Hela，CHO及NIH 3T3细胞的转染效率至少减少50%。Opti-MEMTM可能含有较少的血清，导致转染效率的大幅降低。
4、不含血清的高糖DMEM是很好的选择，可以与其他的生长培养基兼容，如果未能得到较高的转染效率，您可能会考虑更换稀释液，若是您不明确如何选择合适的稀释液，建议您初次可以尝试不含血清的高糖DMEM，您应该会得到满意的转染效率。
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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