HiPure Plasmid EF Mega Kit

Introduction The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture. Details Specifications Features Specifications Main Functions Isolation up to 10mg endotoxin-free plasmid DNA from 500ml bacterial culture Applications Cell transfection, animal injection, etc. Purification method Mega 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: 500ml culture medium Yield 10mg Elution volume ≥0.8ml Time per run ≤70 minutes Liquid carrying volume per column 50ml Binding yield of column 10mg 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 10mg plasmid can be binded in one column Low endotoxin – the obtainedplasmid can be directly used for cell transfection and animal injection, etc. Kit Contents Contents P111602 P111603 Purification Times 10 Preps 50 Preps RNase A 60 mg 2 x 150 mg Buffer E1 220 ml 2 x 550 ml Buffer E2 220 ml 2 x 550 ml Buffer E3 220 ml 2 x 550 ml Buffer E4 220 ml 2 x 550 ml Buffer E5 120 ml 550 ml Buffer PW2 25 ml 2 x 100 ml Elution Buffer 30 ml 120 ml HiPure EF Maxi Columns 10 50 Lysate Clear Maxi Syringe 10 50 50 ml Collection Tubes 20 100 Storage and Stability The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. 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