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NA0500 Sigma

GenElute HP Plasmid Megaprep Kit Green Alternative

sufficient for 5 preparations

Synonym: Gen Elute, GenElute HP Plasmid Kit

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Description

Features and Benefits

• From harvested bacterial culture to pure plasmid DNA in 90 minutes or less
• Up to 5 mg of high-copy plasmid DNA
• Efficient vacuum format
• No phenol/chloroform extraction of alcohol precipitation required

General description

The GenElute HP Plasmid Megaprep Kit offers a simple, rapid, and cost-effective method for isolating plasmid DNA from recombinant E. coli cultures. This high performance kit features vacuum filter units for rapid lysate clearing and for DNA-silica binding. Up to 5 mg of high copy plasmid DNA can be recovered from 200 mL-1.0 L LB medium or 200-600 mL TB medium of E. coli culture in less than 1.5 hours. The actual yield depends on the strain, the plasmid, and the culture medium used.

An overnight recombinant E. coli culture is harvested with centrifugation and subjected to a modified alkaline-SDS lysis procedure followed by adsorption of the DNA onto a silica membrane in the presence of high salts. Contaminants are removed by two wash steps. Finally, the bound DNA is then eluted in Elution Solution (Tris-HCl) or water.

The DNA is ready for immediate use in applications such as restriction digest, cloning, sequencing, PCR, and transfection.

The GenElute HP Plasmid Megaprep Kit offers a simple, rapid, and cost-effective method for isolating plasmid DNA from recombinant E. coli cultures. This high performance kit features vacuum filter units for rapid lysate clearing and for DNA-silica binding. Up to 5 mg of high copy plasmid DNA can be recovered from 200 mL-1.0 L LB medium or 200-600 mL TB medium of E. coli culture in less than 1.5 hours. The actual yield depends on the strain, the plasmid, and the culture medium used.

An overnight recombinant E. coli culture is harvested with centrifugation and subjected to a modified alkaline-SDS lysis procedure followed by adsorption of the DNA onto a silica membrane in the presence of high salts. Contaminants are removed by two wash steps. Finally, the bound DNA is then eluted in Elution Solution (Tris-HCl) or water.

Application

GenElute HP Plasmid Megaprep Kit has been used to purify plasmids.

Sigma Life Science is committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product has Inherently Safer Chemistry, compared to the standard use of phenol and chloroform to perform DNA extractions.

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. The DNA is ready for immediate use in downstream applications such as restriction digestion, ligation, sequencing, PCR, transformation, and transfection.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC

Price and Availability


NAP and qPCR Reagents from Roche

Microcon Centrifugal Filters

All labs need water
Safety & Documentation

Safety Information

Symbol 
Signal word 
Warning
Precautionary statements 
RIDADR 
UN 3316 9
Flash Point(F) 
77 °F
Flash Point(C) 
25 °C

Documents

Certificate of Analysis

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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