如何优化PolyJet转染试剂？
我们使用PolyJet DNA转染试剂转染表皮细胞及Raw267.4细胞非常有效，并且毒性很小。对于如何更好的优化PolyJet转染试剂，我乐意分享以下几点：
1、DNA的质与量。DNA的纯度对于转染实验至关重要。由E Coli制备的DNA，A260/280必须达到1.80甚至更高。对于6孔板，通常每孔~1.0μg DNA足矣。其他规格的细胞培养皿，可以根据表面积适当调整DNA含量。
2、PolyJet/DNA的比率。对于表皮及Raw267.4细胞的转染，我们将PolyJet/DNA的比率固定在3:1，并且得到了满意的结果，没有为寻找更合适的比率问题而烦恼。
3、稀释。DNA及PolyJet的稀释一定不要含有血清。我们使用的是不含血清的高糖DMEM培养基，效果很好。请不要选择Opti-MEM，因为它可以影响转染复合物的形成，因此，千万不要使用Opti-MEM来稀释质粒及PolyJet。
4、转染中可以含有血清。我们在含有血清/抗生素及不含二者的情况下分别进行可转染实验，没有发现两种情况下转染效率有什么差别。因此，血清/抗生素对PolyJet转染试剂没有什么影响。
Some technical tips for optimizing PolyJet reagent.
PolyJet DNA Transfection Reagent is very efficient to transfect epithelial and Raw 267.4 cells in our hands without significant toxicity. Here I would like to share several technical points for better using 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 6-well plate, ~1.0 µg of DNA per well is usually good enough. For other cell culture format, just adjust DNA amount per culture dish's surface area.
2. Ratio of PolyJet/DNA. For epithelial and Raw 267.4 cells, we locked the ratio at 3:1 with excellent results and never bothered to find the optimal ratio.
3. Diluent. Diluent used to dilute DNA and PolyJet reagent must be serum free. Serum-free DMEM medium with high glucose works for us. Opti-MEM is NOT a good choice because it may affect formation of transfection complex. So never use Opti-MEM as dilute.
4. Transfection with serum. We performed transfection with or without
serum/antibiotics and failed to find any difference in efficiency.
Therefore, PolyJet reagent is NOT interfered by serum/antibiotics.
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