HiPure Plasmid EF Midi Kit

Introduction The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water. Details Specifications Features Specifications Main Functions Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture Applications Cell transfection, animal injection, etc. Purification method Midi spin column Purification technology Silica technology Process method Manual (centrifugation or vacuum) Sample type Conventional plasmid, plasmid less than 30KB Sample amount 25-50ml Yield 10-500μg Elution volume ≥0.4ml Time per run ≤60 minutes Liquid carrying volume per column 4ml Binding yield of column 250μg 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 250μg 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 P123102 P123102B Purification Times 50 Preps 50 Preps RNase A 30 mg 30 mg Buffer E1 150 ml 150 ml Buffer E2 150 ml 150 ml Buffer E3 150 ml 150 ml Buffer E4 150 ml 150 ml Buffer E5 150 ml 150 ml Buffer PW2* 50 ml 50 ml Elution Buffer 30 ml 30 ml MaxPure EF Mini Column 50 50 2 ml Collection Tubes 50 50 Lysate Clear Midi Syringe 50 50 Extend Tubes 50 50 50ml Centrifuge Tubes 50 Support Tubes 50 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. After addition of RNase A and optional LyseBlue reagent, 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