The HiPure Plasmid EF Mini Kit combine the power of HiPure technology with Magen's innovative endotoxin removal technology to deliver high-quality plasmid DNA with low endotoxin levels foruse in eukaryotic transfection, and in vitro experiments. The HiPure plasmid endo-free system uses a specially formulated buffer that prevents endotoxin molecules from binding to the surface of the HiPure matrix. Endotoxin contamination lowers transfection efficiencies for endotoxin sensitive celllines. For gene therapy, endotoxin contamination should be of major concernsince endotoxins have the potential to cause fever, endotoxin shock syndrome, and interfere with in vitro transfection into immune cells.

Specifications

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

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，Buffer P1 is stable for 6 months when stored at 2–8°C.