介紹
The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy 質粒DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and 聚合酶鍊式反應 amplification.
細節
規格
| 特徵 | 規格 |
| 主要功能 | Isolation up to 15μg endotoxin free plasmid DNA from 1-5ml bacterial culture |
| 應用領域 | Enzyme digestion, 定序, PCR and labeling, ETC. |
| 淨化技術 | 磁珠技術 |
| 工藝方法 | 手動或自動 |
| 樣品類型 | Conventional plasmid, plasmid≤30KB |
| 樣品量 | 1-5毫升 |
| 洗脫體積 | ≥50μl |
| 每次運行時間 | ≤80 minutes |
原則
This product is based on the purification method of high-binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA 釋放到裂解液中. 加入磁性顆粒和結合溶液後, DNA will be adsorbed on the surface of magnetic particles, 蛋白質等雜質不會被吸附而被去除. The adsorbed particles were washed with a washing solution to remove proteins and impurities, 用乙醇洗滌以除去鹽, and finally, DNA was eluted with an Elution Buffer.
優點
- 高純度 – purified plasmid can be directly used in sequencing, enzyme digestion PCR, ETC.
- 快速地 – it takes only 80 minutes to complete the isolation
- 高產 – up to 15μg plasmid can be bound in one column
- Economic – excellent cost-effectiveness performance
套件內容
| 內容 | P181402 | P181403 | P181404 |
| 淨化時間 | 100 準備工作 | 500 準備工作 | 5000 準備工作 |
| 核糖核酸酶A | 10 毫克 | 30 毫克 | 2 X 160 毫克 |
| Buffer P1 | 30 毫升 | 150 毫升 | 2 X 800 毫升 |
| Buffer P2 | 30 毫升 | 150 毫升 | 2 X 800 毫升 |
| Buffer LEN3 | 20 毫升 | 80 毫升 | 800 毫升 |
| Buffer LN4 | 90 毫升 | 400 毫升 | 4 X 980 毫升 |
| MagPure 顆粒 | 3.5 毫升 | 17 毫升 | 3 X 60 毫升 |
儲存和穩定性
RNase A and MagPure Particles 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) 不影響其性能. 其餘套件組件可在室溫下乾燥保存 (15–25°C) 並且至少穩定 18 在這些條件下幾個月. The entire kit can be stored at 2–8°C, but in this case, buffers should be redissolved before use. 使用時確保所有緩衝液均處於室溫. 如果緩衝液中形成任何沉澱物, 37℃溫熱溶解. After the addition of RNase A, Buffer P1 is stable for 6 months when stored at
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