Roche Tripure RNA Isolation Reagent —RNA纯化的优选

Roche Tripure RNA Isolation Reagent—RNA纯化的优选

从细胞中分离总RNA的纯度与完整性对于许多分子生物学实验至关重要，例如，Northern 印迹及杂交分析，寡聚纤维素选择分离mRNA，cDNA文库的构建等实验的成败在很大程度上取决于总RNA的提取质量。但是做过RNA纯化的人都知道，准备这些样品往往耗时耗力，这主要是因为即稳定又到处可见的RNAses很容易就把这些样品降解。

传统的Trizol方法提取各种样本的RNA，由于步骤繁多，试剂杂乱，往往给研究者带来时间，**以及重复性困惑。这里我们向您推荐引用文献高，操作简便，品质保证的罗氏Tripure Isolation Reagent 试剂盒。

Tripure Isolation Reagent试剂盒的优势：

（1）即用型试剂，提供省时而简单的实验步骤，整个RNA的分离纯化过程只需1-2 小时；

（2）适用各种样本，例如细胞培养液，冰冻切片，人的白细胞，**悬液，酵母以及植物细胞等；

（3）高质量的RNA：A260/A280=1.6-2.0, 可以直接应用于Northern 杂交、体外转录、RNA酶保护分析、cDNA合成以及RT-PCR。

Tripure Isolation Reagent 试剂盒提取原理：

通过一步法裂解样本后，Tripure Isolation Reagent 能够分解细胞并使内源性核酸酶变性。然后通过添加氯仿混合并离心，整个样本被分离成三个相：无色的上层（RNA）、白色的中层和红色的有机底层（DNA和蛋白质）。*后通过乙醇沉淀从而将RNA分离出来。

典型的实验：

Lane1: RNA ladder

Lanes 2 -4: RNA from human leukemia cell line

Lanes 5 -7: RNA from human white blood cell pellet

Lanes 8 -10: RNA from human bloodbuffy coat

Lanes 11 -13: RNA from rat livertissue.

The amount of total RNA applied to the gel was either 5 µg (lanes 2, 5, 8, 11), 1 µg (lanes 3, 6, 9, 12), or 0.25 µg (lanes 4, 7, 10, 13). 

Startingmaterial                                       RNAyield                                DNAyield          

Liver tissue                                               6-10 µg/mg tissue                      3-4 µg/mg tissue

Spleen tissue                                             6-10 µg/mg tissue                       n.d.

Kidney tissue                                            3-4 µg/mg tissue                 3-4 µg/mgtissue Skeletal muscle or braintissue                 1-1.5µg/mgtissue                     2-3 µg/mg tissue

Placenta                                                    1-4 µg/mg tissue                         2-3 µg/mg tissue

Cultured epithelial cells                           8-15 µg/10⁶ cells                         n.d.

Cultured fibroblasts                                 5-7 µg/10⁶ cells                            n.d.

Other cultured human, mouse, or rat cells   n.d.                                         5-7 µg/10⁶ cells

产品订购信息：

 Figure 1: Northern blot of total RNA isolated with the TriPure Isolation Reagent.Total RNA was isolated from the following samples: 1.5 x 10⁶ cells of a human leukemia cell line, 5.0 x 10⁷ human white blood cells, 1.7 x 10⁷ buffy coat cells from human blood, and 500 mg rat tissue. The isolated RNA samples were separated electrophoretically on a gel, transferred to a nylon membrane, and hybridized with a digoxigenin-labeled DNA probe (1 kb) from the glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene. The blot was incubated overnight with DIG/Genius System reagents for chemiluminescent detection, then exposed to X-ray film for 5 min. The G3PDH probe recognizes a 1.35 kb mRNA.