Effectene Transfection Reagent (1 ml)，301425，Qiagen，凯杰

Effectene Transfection Reagent

对原代细胞和敏感细胞系的DNA转染

Features

在血清存在的情况下进行高效转染

只需使用少量DNA 即可达到高效转染

高转染效率而且较低的DNA使用量

适合高通量筛选

Effectene Transfection Reagent是一种非脂质体的脂类转染剂，能将DNA转入多种类型的细胞。因其毒性低，非常适合敏感细胞如原代细胞的转染。

Effectene Transfection Reagent 的操作流程简单, 转染质粒DNA比用其他的试剂获得的转染效率高。Effectene Transfection Reagent适用于带有寡核苷酸的敏感细胞系的转染。特别适用于原代细胞。 使用Effectene Transfection Reagent已成功转染了许多细胞系和原代细胞。有针对不同细胞类型的转染方案。细胞毒性低，因为可在血清存在的情况下使用Effectene Transfection Reagent进行转染且DNA用量低。

DNA重组技术应用于药物研发领域，导致对高通量转染需求增加。使用Effectene Transfection Reagent进行转染DNA用量低，手动操作少。此外，无需移除转染复合物，使该试剂高度适用于高通量筛选。Effectene Transfection Reagent适用于批量转染。

Effectene Transfection Reagent是一种全新的非脂质体的脂类转染剂，将DNA 与浓缩增强子和优化的缓冲液混合使用以达到高效转染。浓缩增强子先聚合DNA分子，Effectene Transfection Reagent随后用阳离子脂质包围聚合的DNA分子。采用一种特殊有效的方法将DNA转入真核细胞。这种特性保证了复杂复合物转染的可重现性。

Effectene流程分为两步。将DNA与浓缩增强子混合后，加入Effectene Transfection Reagent形成复合物，这一步只需2–5分钟。然后加入Effectene Transfection Reagent，与混合物一起反应5–10分钟以形成Effectene–DNA复合物。复合物与培养基混合（可以含有血清和抗体），直接加入到细胞中。培养细胞至成熟，分析基因表达。

High transfection efficiencies using Effectene Reagent.

Transfection efficiencies using Effectene Reagent and two commonly used lipid-based reagents were compared. Murine teratocarcinoma F9 cells (5 x 105) were transfected in 6-well plates with a luciferase-reporter plasmid using optimized conditions based on the manufacturer's instructions for each reagent. Transfection efficiencies were determined by measuring luciferase activity 48 hours post-transfection, and are given as relative light units (RLU). (Data kindly provided by I. Clavereau, D. Petitprez, and I. Van Seuningen, Unité INSERM 377, Place de Verdun, Lille Cedex, France.)

Serum and DNA quantity vs. transfection efficiency.

The influence of serum and DNA quantity on transfection using Effectene Reagent was examined. COS-7 cells (2 x 104 per well) were seeded in 96-well plates one day before transfection. Cells were transfected using 0.01–1.0 µg beta-galactosidase-reporter plasmid and 0.08–8.0 µl Enhancer (DNA: Enhancer ratio of 1:8) and 2 µl Effectene Reagent, in either the presence or absence of serum. Each bar represents the average efficiency from 4 replicates 48 hours post-transfection.

Effectene transfection procedure.

DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2-5 minutes. Effectene Reagent is then added and the mixture is incubated for 5-10 minutes to allow Effectene-DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression.