GenElute™ HP Select Plasmid Gigaprep Kit

Description: The GenElute™ HP Select Plasmid Gigaprep Kit offers a simple, rapid, and cost-effective method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. Up to 15 mg of high copy plasmid DNA with ≤ 0.1 Endotoxin Unit/µg can be recovered from E. coli cultures in 1.2–2.5 liters Luria Broth (LB) medium or 600 ml – 1.2 liters of Terrific Broth (TB) medium in less than 2.0 hours.

• Sophisticated – utilizes HP Select technology, the premier plasmid purification method
• Novel – includes patent-pending endotoxin removal system
• Plentiful – up to 15 mgs of high-quality, endotoxin-free (≤0.1EU/µg) plasmid DNA in 2 hours or less
• Convenient – vacuum format and no ethanol precipitation required

Procedure: An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is cleared by filtration followed by the addition of a binding solution optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured onto a silica membrane in the presence of high salts, while endotoxins are prevented from adsorbing to the membrane. Contaminants are removed with three wash steps. Finally, the bound DNA is eluted in endotoxin-free water.

Application: High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells and gene therapy research.

From harvested bacterial culture to pure plasmid DNA in 2 hours or less

Can be used to purify low-, medium- and high-copy plasmid DNA

Up to 15 mg high-copy plasmid DNA

Vacuum format eliminates need for slow gravity flow columns

Lower cost, higher yields and significantly reduced labor equal substantial savings – in both dollars and time!

Figure 1. Maximum plasmid DNA yield obtained with different constructs using GenElute HP Select Plasmid Gigaprep Kit.

Figure 2. Comparison of transfection efficiencies of high copy plasmid DNA pCMV-SPORT-β-gal/DH5α into endotoxin sensitive cell line HuH-7 cells. The data shows the average and standard deviations of three replicates for the cell line evaluated. Untransfected HuH-7 represent cells treated with transfection reagents without any plasmid DNA.