我們使用PolyJet DNA轉染試劑轉染表皮細胞及Raw267.4細胞非常有效，并且毒性很小。對于如何更好的優化PolyJet轉染試劑，我樂意分享以下幾點：
1、DNA的質與量。DNA的純度對于轉染實驗至關重要。由E Coli制備的DNA，A260/280必須達到1.80甚至更高。對于6孔板，通常每孔~1.0μg DNA足矣。其他規格的細胞培養皿，可以根據表面積適當調整DNA含量。
2、PolyJet/DNA的比率。對于表皮及Raw267.4細胞的轉染，我們將PolyJet/DNA的比率固定在3:1，并且得到了滿意的結果，沒有為尋找更合適的比率問題而煩惱。
3、稀釋。DNA及PolyJet的稀釋一定不要含有血清。我們使用的是不含血清的高糖DMEM培養基，效果很好。請不要選擇Opti-MEM，因為它可以影響轉染復合物的形成，因此，千萬不要使用Opti-MEM來稀釋質粒及PolyJet。
4、轉染中可以含有血清。我們在含有血清/抗生素及不含二者的情況下分別進行可轉染實驗，沒有發現兩種情況下轉染效率有什么差別。因此，血清/抗生素對PolyJet轉染試劑沒有什么影響。
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.