The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-聚合酶鍊式反應, 北方印跡, poly A+ RNA (信使RNA) purification, nuclease protection, and in vitro translation.
細節
規格
| 特徵 | 規格 |
| 主要功能 | Isolation of total RNA from Paxgene RNA Tubes |
| 應用領域 | 逆轉錄聚合酶連鎖反應, 定量PCR, 北方雜交, NGS, nucleic acid protection, in vitro translation |
| 純化方法 | 小型的 旋轉柱 |
| 淨化技術 | 二氧化矽技術, DNA filtration technology |
| 工藝方法 | 手動的 (離心或真空) |
| 樣品類型 | Blood in the preservation tube |
| 樣品量 | 2.5毫升 |
| 洗脫體積 | ≥30微升 |
| 每根柱的載液量 | 800微升 |
| 柱結合率 | 100微克 |
原則
The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanol is added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. The remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.
優點
- 高品質 – high purity total RNA can be directly used in various sensitive downstream applications
- 快速地 – isolation of several samples can be completed in 40 minutes by using the column purification method
- Safety – no phenol-chloroform extraction is required
- 敏感的 – direct lysis of blood, 電漿, and other samples without separation of leukocytes
套件內容
| 內容 | R416802 | R416803 |
| 淨化時間 | 10 準備工作 | 50 準備工作 |
| HiPure RNA Mini Columns I | 10 | 50 |
| gDNA Filter Mini Columns | 10 | 50 |
| 2毫升收集管 | 30 | 150 |
| 無核糖核酸酶水 | 60 毫升 | 250 毫升 |
| Buffer MBR1 | 10 毫升 | 30 毫升 |
| Buffer MBR2 | 5 毫升 | 15 毫升 |
| 緩衝器RW1 | 15 毫升 | 60 毫升 |
| 緩衝器 RW2* | 6 毫升 | 20 毫升 |
| 蛋白酶K | 12 毫克 | 50 毫克 |
| 蛋白酶溶解緩衝液 | 1.8 毫升 | 5 毫升 |
| 脫氧核糖核酸酶I | 120 微升 | 600 微升 |
| DNase Buffer | 6 毫升 | 30 毫升 |
儲存和穩定性
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. 然而, 短期儲存 (DNase I up to 1 週, Proteinase K up to 8 週) 在室溫下 (15–25°C) 不影響他們的表現. 其餘試劑盒成分可在室溫下保存 (15–25°C) 並且至少穩定 18 在這些條件下幾個月.
實驗數據
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