HiPure Plasmid EF Mini Kit

Introduction HiPure Plasmid EF Mini Kit Overview: Technology Integration: Combines HiPure technology with Magen’s innovative Endotoxin Removal Technology (ETR) to ensure high-quality plasmid DNA purification. Endotoxin Management: Features a specially formulated buffer designed to prevent endotoxins from binding to the HiPure matrix, significantly reducing endotoxin levels in the final DNA product. Application Suitability: Provides plasmid DNA ideal for eukaryotic transfection and in vitro experiments, especially where low endotoxin levels are critical. Impact of Endotoxin Contamination: Endotoxin contamination can drastically lower transfection efficiencies, particularly in endotoxin-sensitive cell lines. Clinical Relevance: In contexts such as gene therapy, controlling endotoxin levels is crucial due to the potential for serious adverse effects like fever, endotoxin shock syndrome, and negative impacts on in vitro transfection into immune cells. Details Specifications Features Specifications Main Functions Isolation up to 70µg endotoxin-free plasmid DNA from 10-15ml bacterial culture Applications Cell transfection, animal injection, etc. Purification method Mini spin column Purification technology Silica technology Process method Manual (centrifugation or vacuum) Sample type Conventional plasmid Sample amount High copy plasmid: 10ml culture medium Low copy number plasmid : 10-15ml culture medium Yield 10-70μg Elution volume ≥75μl Time per run ≤40 minutes Liquid carrying volume per column 800μl Binding yield of column 70μg Principle The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA). Advantages High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc. Fast – silica gel column purification is much faster than ion exchange High yield – up to 70μg plasmid can be binded in one column Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection Kit Contents Contents P115402 P115403 Purification Times 50 Preps 250 Preps RNase A 5 mg 20 mg Buffer P1 30 ml 140 ml Buffer P2 30 ml 140 ml Buffer LEN3 15 ml 70 ml Buffer LN4 50 ml 250 ml Buffer LN5 30 ml 140 ml Buffer PW1 30 ml 140 ml Buffer PW2 12 ml 50 ml Elution Buffer 15 ml 30 ml HiPure DNA Mini Columns III 50 250 2 ml Collection Tubes 50 250 Storage and Stability Storage Instructions: General Storage: Kit components can be stored dry at room temperature (15–25°C). Stable for at least 18 months under these conditions. Handling Precipitates: If any precipitates form in the buffers, warm at 37°C to dissolve. Specific Storage for Buffer P1 with RNase A: After addition of RNase A, Buffer P1 is stable for 6 months when stored at 2–8°C. Purchase Guide Name CAT NO Sample amount Column type Elution volume Yield Time per run Centrifuge requirements HiPure Plasmid EF Mini Kit P1154 10-15ml 1.5ml column ≥75μl 10–70µg/15ml ≤40 minutes small centrifuge HiPure Plasmid EF 96 Kit P1157 1-5ml 96 well plate ≥75μl 30µg/1ml ≤60 minutes 96 well plate centrifuge HiPure Plasmid EF Midi Kit P1231 30-50ml 15ml column ≥0.4ml 10-500μg/50ml ≤60 minutes 15ml centrifuge HiPure Plasmid EF Maxi Kit P1156 100-200ml 50ml column ≥0.8ml 100–1500µg/200ml ≤60 minutes 50ml centrifuge HiPure Plasmid EF Mega Kit P1116 500ml 50ml column ≥0.8ml 10mg/500ml ≤70 minutes 50ml centrifuge