HiPure Plasmid EF Maxi Kit

Introduction HiPure Plasmid EF Maxi Kit Features: Technology Integration: The kit integrates HiPure technology for plasmid purification with Magen’s innovative Endotoxin Removal Technology (ETR). Endotoxin Removal: Specifically designed to achieve low levels of endotoxins in the purified plasmid DNA, enhancing its suitability for sensitive applications. Application Suitability: Ideal for use in eukaryotic transfection and various in vitro experiments due to the high purity and low endotoxin levels of the DNA. Capacity and Yield: Capable of purifying up to 1000 µg of high copy number plasmid DNA or 200 µg of low copy number plasmid DNA. Culture Volume Processing: Efficient purification from up to 150 mL of an overnight bacterial culture. Details Specifications Features Specifications Main Functions Isolation of up to 1.5 mg endotoxin-free plasmid DNA from 200ml bacterial culture Applications Cell transfection, animal injection, etc. Purification method Maxi spin column Purification technology Silica technology Process method Manual (centrifugation or vacuum) Sample type Conventional plasmid, large plasmid Sample amount High copy plasmid: 100-150ml culture medium Low copy plasmid: 150-200ml culture medium Yield 100-1500μg Elution volume ≥0.8ml Time per run ≤60 minutes Liquid carrying volume per column 20ml Binding yield of column 1 mg Principle HiPure Plasmid Procedure: Alkaline Lysis and DNA Adsorption: Based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane in the kit replaces glass or silica slurries for plasmid DNA minipreps. Three Basic Steps: Step 1: Preparation and clearing of a bacterial lysate using the alkaline method. Step 2: Transfer of the supernatant to a column for DNA binding. Step 3: Washing to remove proteins and other impurities, followed by elution of nucleic acid with low-salt buffer (10 mm Tris, pH 9.0, 0.5 mm 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 1.5 mg plasmid can be binded in one column Ultra low endotoxin — content of endotoxin is less than 0.05 EU/μg, which can be directly used for cell transfection and animal injection, etc. Kit Contents Contents P115602 P115603 Purification Times 10 Preps 50 Preps RNase A 30 mg 150 mg Buffer P1 100 ml 500 ml Buffer P2 100 ml 500 ml Buffer LN3 50 ml 250 ml Buffer PW1 33 ml 180 ml Buffer PW2 20 ml 100 ml Elution Buffer 15 ml 120 ml Buffer CL 33 ml 180 ml HiPure DNA Maxi Columns C 10 50 Lysate Clear Midi Syringe 10 50 50 ml Collection Tubes, C 20 100 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. Buffer Precipitates: If any precipitates form in the buffers, warm at 37°C to dissolve. Specific Storage for RNase A and Buffer P1: RNase A and Buffer P1 are 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