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NA0410 Sigma-Aldrich

GenElute HP Endotoxin-Free Plasmid Maxiprep Kit

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sufficient for 25 preparations

Synonym: Gen Elute, GenElute Endotoxin-Free Plasmid Kit

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Description

Features and Benefits

• Up to 1.2 mg of high-copy plasmid DNA with endotoxin levels of ≤0.1 EU/μg
• Only 35 minutes from pelleted cells to purified plasmid
• Convenient vacuum format

General description

The GenElute HP Endotoxin-Free Plasmid Maxiprep Kit offers a simple and rapid method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. The kit uses a vacuum format with a filter column for the rapid clearing of the bacterial lysate and a silica column for capturing plasmid DNA. A proprietary solution binds plasmid DNA to the binding column while preventing endotoxins from adsorbing. The technology allows for the user to consistently achieve levels of endotoxin that are less than 0.1 endotoxin units per μg of plasmid DNA.

High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells.

Other Notes

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

Principle

An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is clarified by filtration followed by the addition of a binding solution that has been optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured on silica, while endotoxins are prevented from adsorbing to the membrane. Two wash steps remove contaminants. Finally, the bound DNA is eluted in endotoxin-free water. The recovered plasmid DNA is predominately in its supercoiled form. Genomic DNA and RNA are below detectable levels by ethidium bromide stained agarose gel electrophoresis.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

Symbol 
GHS06  GHS06
Signal word 
Danger
Hazard statements 
Supplemental Hazard Statements 
Contact with acids liberates very toxic gas.
RIDADR 
UN 2924 8(3) / PGIII

Milli-Q® Water Purification Solutions
Protocols & Articles

Articles

Plasmid DNA Purification Overview

Extraction of macromolecules such as DNA, RNA, and protein is one of the basic methods used in molecular biology. The process of extraction and purification of nucleic acids has evolved from being a ...
Keywords: Centrifugation, Chromatography, Cloning, DNA purification, Digestions, Genetic, Ion Exchange, Molecular biology, Nucleic acid denaturation, Nucleic acid purification, Polymerase chain reaction, Precipitation, Purification, Sequencing, Transcription, transformation

Peer-Reviewed Papers
15

References

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