GenElute™ HP Select Plasmid Gigaprep Kit

Description

• General description: The GenElute HP Select Plasmid Gigaprep Kit offers a simple, rapid, and cost effective method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. Up to 15 mg of high copy plasmid DNA with ≤0.1 endotoxin unit/μg plasmid DNA can be recovered from E. coli cultures in 1.2-2.5 L Luria Broth (LB) medium or 600 mL-1.2 L of Terrific Broth (TB) medium in less than 2 hours after harvesting the cells. Note that the actual yield depends on the strain, the plasmid, and the culture medium used.An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is cleared by filtration followed by the addition of a binding solution optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured onto a silica membrane in the presence of high salts while endotoxins are prevented from adsorbing to the membrane. Contaminants are removed with three wash steps. Finally, the bound DNA is eluted in endotoxin-free water.The DNA is ready for immediate use in applications such as restriction digest, cloning, sequencing, PCR, and transfection of endotoxin-sensitive cell lines.
• Application: GenElute® HP Select Plasmid Gigaprep Kit has been used to purify plasmids.[1]
• Features and Benefits: Up to 15 mg with endotoxin levels of ≤ 0.1 EU/μgOnly 90 minutes from pelleted cells to purified plasmidVacuum format with no ethanol precipitationCan purify low-, medium-, and high-copy plasmid DNA
• Other Notes: For additional information, please see www.sigma-aldrich.com/genelutehp.
• Legal Information: Gabitril is a registered trademark of Abbott Laboratories Corp.