介紹
- 快速地, Simple, and Cost-Effective: Offers a rapid, easy-to-use, and economical method for 質粒DNA miniprep suitable for routine molecular biology applications.
- Silica Membrane Technology: Utilizes silica membrane technology, eliminating the need for cumbersome steps associated with loose resins or slurries, streamlining the purification process.
- Immediate Usability: Purified plasmid DNA is immediately ready for downstream applications, saving time and effort.
- No Phenol Extraction or Ethanol Precipitation: Bypasses the need for phenol extraction and ethanol precipitation, simplifying the workflow and reducing hands-on time.
- High-Quality DNA: Yields high-quality plasmid DNA eluted in a small volume of Tris buffer or water, ensuring optimal performance in downstream applications.
規格
特徵 | 規格 |
主要功能 | Isolation of up to 35μg plasmid DNA from 1-5ml bacterial culture |
應用領域 | Enzyme digestion, 定序, 聚合酶鍊式反應, cloning, ETC. |
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 | Conventional plasmid, plasmid less than 30KB |
樣品量 | High copy plasmid: 1-5ml culture medium Low copy number plasmid: 5-10ml culture medium |
屈服 | 5-35微克 |
洗脫體積 | ≥30μl |
每次運行時間 | Complete 1-24 samples in 30 分分鐘 |
每根柱的載液量 | 800微升 |
柱結合率 | 35微克 |
原則
- Alkaline Lysis: Begins with the alkaline lysis of bacterial cells, which breaks down the cell membrane and releases plasmid DNA into the lysate.
- Silica-Based DNA Binding: The released DNA is adsorbed onto the silica membrane in the presence of high salt, facilitating efficient binding of DNA to the membrane surface.
- Three Basic Steps: The procedure involves three main steps: (1) Preparation and clearing of the bacterial lysate using an alkaline method, (2) Transfer of the cleared lysate to a column for DNA binding, 和 (3) Washing to remove proteins and other impurities.
- Silica Membrane: Utilizes a unique silica membrane in the kit, which completely replaces traditional glass or silica slurries typically used in plasmid DNA minipreps.
- Elution with Low-Salt Buffer: After washing to remove impurities, 核酸, including plasmid DNA, is eluted from the silica membrane using a low-salt buffer containing 10mM Tris, 酸鹼度 9.0, and 0.5mM EDTA.
優點
- 高純度 – purified plasmid can be directly used in sequencing, enzyme digestion PCR, ETC.
- 快速地 – it takes only 1 minute to obtain supernatant by optimized solution (10 minutes for other brands)
- 高產 – up to 35µg plasmid can be bound in one column
套件內容
內容 | P100102 | P100103 |
淨化時間 | 100 準備工作 | 250 準備工作 |
核糖核酸酶A | 5 毫克 | 10 毫克 |
Buffer P1 | 30 毫升 | 80 毫升 |
Buffer P2 | 30 毫升 | 80 毫升 |
Buffer P3 | 40 毫升 | 100 毫升 |
Buffer PW1 | 60 毫升 | 140 毫升 |
Buffer PW2* | 20 毫升 | 50 毫升 |
洗脫緩衝液 | 15 毫升 | 30 毫升 |
HiPure DNA Mini Columns II | 100 | 250 |
2 毫升收集管 | 100 | 250 |
儲存和穩定性
- 儲存條件: Kit components can be stored dry at room temperature (15-25℃) and remain stable for a minimum of 18 在這些條件下幾個月.
- Buffer Precipitates: In case of any precipitates forming in the buffers, warm them at 37°C to dissolve effectively.
- Buffer P1 Stability: After the addition of RNase A, Buffer P1 remains stable for up to 6 months when stored at 2-8°C.
實驗數據
Purchase Guide
姓名 | CAT NO | 樣品量 | Column type | 洗脫體積 | 屈服 | 每次運行時間 | Centrifuge requirements |
HiPure 質粒迷你試劑盒 | P1001 | 1-5毫升 | 1.5ml column | 30-50微升 | 5–35µg/5ml | ≤30 minutes | small centrifuge |
HiPure Plasmid Mini Kit II | P1002 | 5-15毫升 | 1.5ml column | 75-100微升 | 20-80µg/15ml | ≤30 minutes | small centrifuge |
HiPure Plasmid Plus 96 成套工具 | P1006 | 1-1.5毫升 | 96 孔板 | 70-150微升 | 1-15μg/1ml | ≤60分鐘 | 96-well plate centrifuge |
HiPure Fastfilter Plasmid Midi Kit | P1013 | 30-50毫升 | 15ml column | 0.3-0.8毫升 | 50–250µg/50ml | ≤60分鐘 | 15ml centrifuge |
HiPure Fastfilter Plasmid Maxi Kit | P1014 | 100-200毫升 | 50ml column | 0.5-1.5毫升 | 0.4–1mg/200ml | ≤60分鐘 | 50ml centrifuge |
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