快速简单的DNA 转染，用于标准细胞株

Features

优化的细胞特异性转染方案

操作简便迅速

细胞存活率高，毒性低

Product Details

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。

Performance

PolyFect Transfection Reagent保持高效转染率。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。

Principle

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。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，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。

Procedure

PolyFect Transfection Reagent是即用型试剂。转染流程简便迅速，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。

高通量转染

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。

Applications

PolyFect Transfection Reagent特别适用于标准细胞系高效转染，主要用于：

基因表达与功能研究

药物研究与开发

Supporting data and figures

High transfection efficiencies in CHO and 293 cells.

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.

Specifications

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 Up to 50–100 transfections in 6-well plates / 1 ml reagent

Nucleic acid                                     DNA

Cell type                                          COS-7, NIH/3T3, HeLa, 293, CHO cells