usage
sufficient for 15 purifications
Quality Segment
sustainability
Sustainability features
technique(s)
DNA purification: suitable
test parameters
: <2 hr kit run time for up to 12 preps, sample volume: 25-200 mL bacterial culture
storage temp.
15-25°C
General description
The GenElute Endotoxin-free Plasmid Kit offers a simple, rapid, cost effective method for purifying plasmid DNA with ≤0.1 EU/μg DNA for high efficiency transfection. Endotoxins (also known as lipopolysaccharides or LPS) are often co-purified with plasmid DNA and significantly reduce transfection efficiencies in endotoxin-sensitive cell lines.
Recombinant E. coli is harvested from an overnight culture by centrifugation and subjected to a modified alkaline-SDS lysis procedure to produce a cleared lysate. Endotoxins are removed from the cleared lysate with simple extraction-phase separation steps. Plasmid DNA is further purified by adsorption onto silica in the presence of high salts. After a simple spin-wash step, the bound plasmid DNA is eluted in endotoxin-free water. Up to 250 ug of endotoxin-free plasmid DNA may be prepared from 5-40 mL of culture.
The recovered plasmid DNA is predominately in its supercoiled form. The DNA is ready for immediate use in downstream applications such as transfection, restriction digestion, cloning, sequencing, and PCR amplification.
Recombinant E. coli is harvested from an overnight culture by centrifugation and subjected to a modified alkaline-SDS lysis procedure to produce a cleared lysate. Endotoxins are removed from the cleared lysate with simple extraction-phase separation steps. Plasmid DNA is further purified by adsorption onto silica in the presence of high salts. After a simple spin-wash step, the bound plasmid DNA is eluted in endotoxin-free water. Up to 250 ug of endotoxin-free plasmid DNA may be prepared from 5-40 mL of culture.
The recovered plasmid DNA is predominately in its supercoiled form. The DNA is ready for immediate use in downstream applications such as transfection, restriction digestion, cloning, sequencing, and PCR amplification.
This product aligns with one or more of the 12 Principles of Green ChemistryThis product has Inherently Safer Chemistry, compared to the standard use of phenol and chloroform to perform DNA extractions.
Application
The kit has been used to purify plasmid DNA for high efficiency transfection.
Features and Benefits
- High transfection efficiency and yield
- Purify up to 250 μg endotoxin-free plasmid DNA (≤0.1 EU/μg DNA)
- Fast and simple allowing up to 12 preps in less than 2 hours
Legal Information
GenElute is a trademark of Sigma-Aldrich Co. LLC
경고어
Danger
Hazard Classifications
Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Eye Irrit. 2 - Met. Corr. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
표적 장기
Central nervous system
인화점 (F)
Not applicable
인화점 (°C)
Not applicable
WGK
WGK 3
저장 등급
10 - Combustible liquids


