HiPure Plasmid Mini Kit

Introduction Fast, Simple, and Cost-Effective: Offers a rapid, easy-to-use, and economical method for plasmid 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. Specifications Features Specifications Main Functions Isolation of up to 35μg plasmid DNA from 1-5ml bacterial culture Applications Enzyme digestion, sequencing, PCR, cloning, etc. Purification method Mini spin column Purification technology Silica technology Process method Manual (centrifugation or vacuum) Sample type Conventional plasmid, plasmid less than 30KB Sample amount High copy plasmid: 1-5ml culture medium Low copy number plasmid: 5-10ml culture medium Yield 5-35µg Elution volume ≥30μl Time per run Complete 1-24 samples in 30 minutes Liquid carrying volume per column 800µl Binding yield of column 35µg Principle 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, and (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, nucleic acid, including plasmid DNA, is eluted from the silica membrane using a low-salt buffer containing 10mM Tris, pH 9.0, and 0.5mM EDTA. Advantages High purity – purified plasmid can be directly used in sequencing, enzyme digestion PCR, etc. Fast – it takes only 1 minute to obtain supernatant by optimized solution (10 minutes for other brands) High yield – up to 35µg plasmid can be bound in one column Kit Contents Contents P100102 P100103 Purification Times 100 Preps 250 Preps RNase A 5 mg 10 mg Buffer P1 30 ml 80 ml Buffer P2 30 ml 80 ml Buffer P3 40 ml 100 ml Buffer PW1 60 ml 140 ml Buffer PW2* 20 ml 50 ml Elution Buffer 15 ml 30  ml HiPure DNA Mini Columns II 100 250 2 ml Collection Tubes 100 250 Storage and Stability Storage Conditions: Kit components can be stored dry at room temperature (15-25°C) and remain stable for a minimum of 18 months under these conditions. 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. Experiment Data Purchase Guide Name CAT NO Sample amount Column type Elution volume Yield Time per run Centrifuge requirements HiPure Plasmid Mini Kit P1001 1-5ml 1.5ml column 30-50μl 5–35µg/5ml ≤30 minutes small centrifuge HiPure Plasmid Mini Kit II P1002 5-15ml 1.5ml column 75-100μl 20-80µg/15ml ≤30 minutes small centrifuge HiPure Plasmid Plus 96 Kit P1006 1-1.5ml 96 well plate 70-150μl 1-15μg/1ml ≤60 minutes 96-well plate centrifuge HiPure Fastfilter Plasmid Midi Kit P1013 30-50ml 15ml column 0.3-0.8ml 50–250µg/50ml ≤60 minutes 15ml centrifuge HiPure Fastfilter Plasmid Maxi Kit P1014 100-200ml 50ml column 0.5-1.5ml 0.4–1mg/200ml ≤60 minutes 50ml centrifuge