介紹
Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 分分鐘. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, 電漿, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-聚合酶鍊式反應, Northern hybridization and virus detection.
細節
規格
特徵 | 規格 |
主要功能 | Extract viral RNA from 140μl cell-free samples |
應用領域 | 逆轉錄聚合酶連鎖反應, 北方雜交, and various virus detection |
純化方法 | 小型的 旋轉柱 |
淨化技術 | 二氧化矽技術 |
工藝方法 | 手動的 (離心或真空) |
樣品類型 | Cell-free body fluid or culture medium |
樣品量 | 140微升 |
洗脫體積 | ≥15μ |
每次運行時間 | ≤25分鐘 |
每根柱的載液量 | 800微升 |
柱結合率 | 100微克 |
原則
Hipure矽膠柱以高結合力為基礎的玻璃纖維濾膜. 高濃度電離劑條件下 (例如 胍 鹽酸鹽或鳥選擇異硫氰酸鹽), 濾膜可透過氫鍵和靜電吸附核酸, 雖然蛋白質和其他雜質沒有吸附和去除. 清洗吸附有核酸的濾膜,去除殘留的蛋白質和鹽分. 最後, 吸附在濾膜上的核酸可以用低鹽緩衝液洗掉 (例如緩衝液TE) 或水. 所得的核酸純度高,可直接用於各種下游實驗.
The sample is homogenized and lysed in the lysate, and RNA is released into the lysate. The high concentration of guanidine isothiocyanate contained in the lysate denatured and inactivated endogenous or exogenous RNase, while RNA is protected from degradation. The lysate is centrifuged to remove insoluble impurities. After adding ethanol to adjust the binding conditions, it is transferred to the column for filtration. RNA is adsorbed on the membrane of the column, while protein is removed without adsorption. The column is washed with buffer VHB to remove protein and other impurities, washed with buffer RW2 to remove salt. Finally the RNA is eluted by RNase free water. The eluted RNA can be directly used in RT-PCR, 北印跡, poly-A purification, 體外翻譯, ETC.
優點
- 快速地 – 可以提取多個樣本 20 分鐘柱法
- 高品質 – 高純度總RNA可直接用於各種敏感下游應用
- 安全的 – 無需苯酚氯仿萃取
- 敏感的 – RNA can be recovered at the level of PG
- 高產 – carrier RNA contained in the product maximize the recovery of trace nucleic acid
套件內容
內容 | R417102 | R417103 |
淨化時間 | 50 準備工作 | 250 準備工作 |
HiPure Viral Micro Columns | 50 | 250 |
2毫升收集管 | 100 | 500 |
Buffer VRL | 50 毫升 | 200 毫升 |
載體RNA | 310 微克 | 3 X 310 微克 |
Buffer VHB* | 13 毫升 | 110 毫升 |
緩衝器 RW2* | 20 毫升 | 2 X 50 毫升 |
無核酸酶水 | 10 毫升 | 30 毫升 |
儲存和穩定性
Carrier RNA should be stored at 2–8°C upon arrival. 然而, 短期儲存 (取決於 12 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 months under theseconditions
評論
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