GeneJET Endo-Free Plasmid Maxiprep Kit

Thermo Scientific GeneJET Endo-Free Plasmid Maxiprep Kit is a fast, convenient, and efficient system for the large scale purification of endotoxin-free plasmid DNA. Endotoxins are major contaminants found in plasmid DNA preparations. For transfection of sensitive cell lines endotoxins are a major problem. The same applies to in vivo transfection as endotoxins can elicit strong immune responses. Thermo Scientific GeneJET Endo-Free Plasmid Maxiprep Kit is designed to reduce endotoxin level to <0.1 EU/μg of plasmid DNA. The kit utilizes a silica-based membrane technology in the form of convenient spin columns.

Each preparation recovers up to 1000 μg of high copy plasmid DNA from 350 mL of recombinant E. coli cultures that can be used in a wide variety of molecular biology procedures such as transfection of sensitive cells (e.g. Huh-7), in vivo transfection, transformation, restriction endonuclease digestion, automated sequencing, and in vitro transcription.

Highlights

• Pure—free of endotoxins plasmid DNA suitable for transfection of sensitive cells
• Efficient—plasmid recovery resulting in high DNA yields
• Fast—isolation of high quality plasmid DNA in less than 60 minutes
• Convenient—spin column format, centrifuge or vacuum protocol

Applications

Isolated plasmid DNA is suitable for all conventional molecular biology procedures, including:

• Transfection of sensitive cell lines
• FastDigest restriction enzyme digestion
• In vivo transfection
• Automated fluorescent sequencing
• PCR
• Cloning
• Transformation
• In vitro transcription

Includes

• Resuspension Solution
• Lysis Solution
• Neutralization Solution
• Endotoxin Binding Reagent
• Wash Solution I (concentrated)
• Wash Solution II (concentrated)
• RNase A Solution
• Elution Buffer (10 mM Tris-HCl, pH 8.5)
• GeneJET Endo-Free Maxi Purification Columns & Collection Tubes (50 mL, blue-capped)
• GeneJET Endo-Free Maxi Filtration Columns & Collection Tubes (50 mL, white-capped)

Hazardous: No

Quality Control:

• The GeneJET Plasmid Maxiprep Kit is qualified by isolating high copy number plasmid DNA from 250 mL of overnight E. coli culture (OD600 = 2) grown in LB medium following the protocol outlined in the manual.
• The kit passes QC requirements if the purified plasmid DNA has an A_260/280 ratio between 1.8 and 1.9 and a dominant band of supercoiled plasmid DNA is observed after agarose gel electrophoresis.
• The functional quality of the plasmid DNA is evaluated by digestion with restriction endonucleases.