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PolyFect Transfection Reagent采用先进的活化树状多聚物,从多聚物中心向外辐射出带正电荷的氨基基团与DNA 分子结合,为下列细胞株特别优化:COS-7、NIH/3T3、HeLa、293、CHO。
PolyFect Transfection Reagent保持高效转染率(参见" High transfection efficiencies in CHO and 293 cells"和" PolyFect Reagent with HeLa cells")。与其他试剂不同的是,PolyFect Transfection Reagent能够在血清存在的情况下进行转染而不降低转染效率。
PolyFect Transfection Reagent是一种特殊活化的树状聚合物。它由从中心发散出分支的球形大分子构成(参见" Activated dendrimer")。分支终止于带电的氨基,可与带相反电荷的磷酸基相互作用。PolyFect Reagent将DNA聚集成紧密结构(参见 " PolyFect–DNA interaction"),附于细胞表面通过非特异性内吞作用进入细胞。该试剂调节细胞核内pH,抑制核内核酸,从而保证了PolyFect–DNA复合物的稳定性。
PolyFect Transfection Reagent是即用型试剂。转染流程(参见" PolyFect transfection procedure")简便迅速,只需要将试剂加入到DNA溶液中混合,反应5–10分钟,加入培养基(可以含血清和抗体),将PolyFect–DNA复合物移到细胞上。培养细胞用于基因表达。无需转染后移除复合物或更换及增加培养基,操作简便快速。
使用PolyFect Transfection Reagent进行转染只需少量手动操作且无需移除转染复合物,特别适用于高通量筛选。PolyFect Reagent在高通量转染中转染效率高,可重复性好,细胞毒性低,适用于批量转染。关于在96孔板中对COS-7、NIH/3T3、HeLa、HeLa-S3、293和CHO细胞进行转染的实验方案,请咨询QIAGEN技术支持。
PolyFect Transfection Reagent特别适用于标准细胞系高效转染,主要用于:
Transfection efficiencies for PolyFect Reagent and 2 lipid-based reagents were compared. Cells cultured in 6-well plates were transfected with a beta-galactosidase-reporter plasmid. The appropriate protocol was used for transfection with PolyFect Reagent. Optimized protocols were used for transfection with Reagents L2 and F; these were: 10 µl Reagent L2 with 3 µg DNA and 15 µl Reagent F with 2 µg DNA for CHO cells, and 10 µl Reagent L2 with 2 µg DNA and 7 µl Reagent F with 3 µg DNA for 293 cells. All transfections were performed in triplicate. Two independent transfection experiments are shown for 293 cells.
Features | Specifications |
---|---|
Applications | RNAi studies, gene expression studies... |
Transfection type | Transient transfection, stable transfection, plasmid co-transfection |
Controls | Not included |
Features | Optimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum |
Technology | Activated dendrimers |
Number of possible transfections | 25–10,000 transfections |
Nucleic acid | DNA |
Cell type | COS-7, NIH/3T3, HeLa, 293, CHO cells |