GenElute^™ Five-Minute Plasmid Miniprep Kit

Application

The GenElute Five-Minute Plasmid Miniprep Kit is intended for rapid preparation of plasmid DNA for use in capillary DNA sequencing, clone screening, restriction digestion, and PCR. For low copy plasmids, the kit can be used to prepare plasmid DNA for use in restriction digestion and PCR.

Features and Benefits

• No pelleting cells or clearing lysates
• Binding column works with any standard laboratory vacuum manifold
• Up to 5 μg of plasmid DNA or enough for 15 sequencing reactions
• DNA suitable for immediate capillary DNA sequencing

Other Notes

For additional information, please see www.sigma-aldrich.com/genelutehp.

General description

The GenElute Five-Minute Plasmid Miniprep Kit features an ultra-streamlined protocol resulting in high-quality plasmid DNA in about five minutes.

Recombinant E. coli overnight culture in LB (Luria-Bertani) broth is treated briefly (1-2 minutes) with a lysis reagent to effect rapid cell lysis and RNA degradation, without prior removal of culture medium. A binding solution is then mixed with the lysate, and DNA is captured on a silica-based binding column. Impurities are removed by a wash solution and bound plasmid DNA is then eluted in Tris buffer or water, ready for immediate use in various downstream applications. Typical yields of high copy plasmids are 2-6 mg from 400 mL of overnight culture in LB broth.

The GenElute Five-Minute Plasmid Miniprep Kits utilize a novel lysis procedure that eliminates the need to pellet cells from an overnight culture, remove growth media or clear a neutralized lysate. 400 μl of culture is added to a 2 mL collection tube and cells are lysed for 1−2 minutes. A Binding Solution is then added and the entire mixture is passed through a binding column. After washing the column to remove trace contaminants, the plasmid DNA is eluted in 10 mM Tris HCl, pH 8.5. Molecular biology water may be substituted as an elution reagent.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC