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PFM250 Sigma-Aldrich

GenElute Five-Minute Plasmid Miniprep Kit

sufficient for 250 purifications

Synonym: Gen Elute, GenElute Plasmid Miniprep Kit

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Description

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 μg from 400 μL 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

Safety & Documentation

Safety Information

Symbol 
Signal word 
Danger
Precautionary statements 
Supplemental Hazard Statements 
Contact with acids liberates very toxic gas.
RIDADR 
UN 3316 9
Flash Point(F) 
69.8 °F
Flash Point(C) 
21 °C

Milli-Q® Water Purification Solutions
Protocols & Articles

Articles

Blue-White Screening & Protocols for Colony Selection

Blue-white screening is a rapid and efficient technique for the identification of recombinant bacteria. It relies on the activity of β-galactosidase, an enzyme occurring in E. coli, which cleaves lac...
Keywords: Antibiotics, Cloning, Culture media, Gene expression, Immunocytochemistry, Molecular biology, Peptide synthesis, Polymerase chain reaction, Recombination, Sterilizations, transformation

Related Content

GenElute™ Five-Minute Plasmid Miniprep Procedure [VIDEO]

The GenElute Five-Minute Plasmid Miniprep kit features an ultra streamlined protocol yielding high-quality plasmid DNA in about five minutes. The DNA is suitable for immediate use in capillary DNA se...
Keywords: Digestions, Polymerase chain reaction, Sequencing

Peer-Reviewed Papers
15

References

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