Basis Strategien a Methoden fir De-Endotoxiniséierung wärend der Plasmid DNA Reinigung

The use of endotoxin-free plasmids should be considered when the extracted plasmid DNA is used for transfection of primary, Suspensioun, oder sensibel Zellen, Gene Silencing Studien, Zellinjektioun Mikromanipulatioun, gene therapy or DNA vaccines,and other extremely demanding experiments.

Plasmid de-endotoxinization refers to the step of removing endotoxin (lipopolysaccharide, LPS) during the purification of plasmid DNA. Endotoxin is a lipopolysaccharide and protein complex in the cell wall of Gram-negative bacteria that is released when the bacteria lyse. The presence of endotoxin greatly affects the transfection efficiency and may activate non-specific reactions of immune cells, resulting in false positives in the experiment. Dofir, in order to ensure a high transfection rate and high survival of transfected cells, endotoxin must be removed from the process of plasmid preparation.

The importance of endotoxin removal is:

1.Affect the transfection efficiency: endotoxin will reduce the transfection efficiency of primary cells and sensitive cultured cells, and interfere with the in vitro transfection of immune cells.

2.Activate immune response: endotoxin can also lead to fever, hypotension, respiratory distress, intravascular coagulation, and endotoxic shock syndrome, endotoxin can stimulate the synthesis of inflammatory factors IL-1, IL-6, IL-10, and TNF-α by activating TLR4.

3,.Drug regulatory requirements: The State Drug Administration stipulates that the content of endotoxin in DNA vaccine products should not be higher than 0.01 EU/μg, and the dose for personal use should not exceed 20 EU.

There are two strategies for endotoxin removal:

1. Removal at the binding stage: endotoxin is removed at the plasmid DNA binding stage.

2. Removal after elution: remove endotoxin after plasmid DNA elution.

The current endotoxin removal methods mainly include:

1. Non-selective removal: such as adsorption, ultrafiltration, and exclusion chromatography.

2. Selective precipitation: z.B., using polymyxin B, histidines, and poly-L-lysine affinity chromatography.

3. Hydrophobic interaction chromatography and ion exchange chromatography.

Eis endotoxin-free plasmid DNA extraction kit (magnetesch Perle Method) adopts a unique solution formula that requires only a few simple washes to remove endotoxin, Proteinen, polysaccharides, an aner Gëftstoffer, and then obtains high-quality plasmid DNA. This product extracts and purifies plasmid DNA with little damage to plasmids, high extraction concentration, and a high proportion of super-helix. The extracted plasmid DNA can be used for digestion, PCR, sequencing, ligation,n and transformation.

Fournisseur

Shanghai Lingjun Biotechnology Co., Ltd., Ltd.gouf etabléiert an 2016 deen e professionnelle Hiersteller vu biomagnetesche Materialien an Nukleinsäure Extraktiounsreagenz ass.

Mir hu räich Erfahrung an Nukleinsäure Extraktioun an Offäll, Protein Reinigung, Zell Trennung, Chemilumineszenz an aner technesch Felder.

Eis Produkter gi wäit a ville Beräicher benotzt, wéi medizinesch Tester, genetesch Tester, Universitéit Fuerschung, genetesch Zucht, an esou weider. Mir bidden net nëmmen Produkter, awer och kënnen OEM ënnerhuelen, ODM, an aner Besoinen.Wann Dir e verbonne Besoin hutt, weg fillen gratis eis ze kontaktéieren umsales01@lingjunbio.com.

• Optimiséierungstemperatur an Zäit fir Silica-baséiert Magnéitpärelen an DNA Fragment Selektioun
• Kënnt Dir ëmmer nach ofgelaf Bio-Nano Magnéitpärelen benotzen?
• Gentle Ëmgank: Non-negotiable Baseline fir DNA Fragment Auswiel
• Präzisioun “Redaktioun” vum Liewen Code: Den Uwendungswäert an technesche Choixe vun der Selektioun vun DNA Fragmenter
• Wéi kann den inhibitoreschen Effekt vu magnetesche Perlen op PCR reduzéiert ginn?