Qiagen 74134/74136 RNeasy Plus Mini Kit 组织细胞总RNA提取试剂盒
For purification of up to 100 µg total RNA from cells/tissues using gDNA Eliminator columns
Cells and easy-to-lyse tissues are lysed and homogenized in highly denaturing guanidine-isothiocyanate–containing Buffer RLT Plus, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate is then passed through a gDNA Eliminator spin column. This column, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the sample is applied to an RNeasy MinElute spin column. These specialized columns contain a silica membrane that specifically binds RNA from lysed cells.
Qiagen 74134/74136 RNeasy Plus Mini Kit 组织细胞总RNA提取试剂盒Total RNA is purified from up to 107 cells or 30 mg tissue. A short workflow enables RNA purification with genomic DNA removal in less than 25 minutes (see flowchart "RNeasy Plus procedure"). Samples are first lysed and homogenized. The lysate is passed through a gDNA Eliminator spin column, ethanol is added to the flow-through, and the sample is applied to an RNeasy MinElute spin column. RNA binds to the membrane and contaminants are washed away. High quality RNA is eluted in 30 µl, or more, of water.
Different protocols are available for different starting materials. The protocols differ mainly in the lysis and homogenization of the sample. Once the sample is applied to the gDNA Eliminator spin column, the protocols are similar. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells.
When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the RNeasy Plus Mini Kit), excessive foaming may occur. This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus before starting disruption and homogenization. Reagent DX has been carefully tested with the kit, and has no effect on RNA purity or on downstream applications.
RNA purified using the RNeasy Plus Mini Kit is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative real-time RT-PCR. The purified RNA can also be used in other applications.
| Features | Specifications |
| 应用 | PCR, qPCR, real-time RT-PCR, microarray |
| 洗脱体积 | 30-50 µl |
| 规格 | Spin column |
| 整合基因组DNA去除步骤 | Yes |
| 主要样本类型 | Tissue, cells |
| 处理 | Manual (centrifugation) |
| 纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 | Total RNA |
| 样本量 | 30 mg (<700 µl) |
| 技术 | Silica technology |
| 每次运行或制备时间 | 25 minutes |
| 产量 | Varies |
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