Maxiprep endotoxin-free plasmid DNA purification—NucleoBond PC 500 EF
NucleoBond PC EF Kits use anion exchange and gravity flow for purification of endotoxin-free plasmid DNA. Endotoxin removal is achieved in a unique washing step—no extra incubations are necessary. Lysates are cleared by simple filtration using the included NucleoBond Folded Filters; no centrifugation is required. NucleoBond PC 500 EF is used to purify 100–500 µg of endotoxin-free plasmid DNA.
NucleoBond PC EF Kits use anion exchange and gravity flow for purification of endotoxin-free plasmid DNA. Endotoxin removal is achieved in a unique washing step—no extra incubations are necessary. Lysates are cleared by simple filtration using the included NucleoBond Folded Filters, no centrifugation is required. NucleoBond PC 500 EF is used to purify 100–500 µg of endotoxin-free plasmid DNA.
Endotoxins (lipopolysaccharides, LPS) are a major component of the Gram-negative bacterial cell wall. The LPS molecule is an extremely potent stimulator of the mammalian immune system, and a number of mechanisms exist to detect LPS and to respond to the presence of either this molecule or Gram-negative bacteria. LPS is a common contaminant of plasmid DNA preparations grown in E. coli. The negative charges associated with lipid A and the inner core of LPS cause the LPS molecule to behave like DNA on anion exchange chromatography resins. An important prerequisite for transfections is endotoxin-free plasmid DNA.
The level of endotoxin contamination in purified plasmid DNA is critical for many applications. For this reason Macherey-Nagel developed a specific patented procedure that reduces endotoxins to a very low level. In comparison to competitor kits, there is no need for a time-consuming incubation step to remove the endotoxins (see User Manual). The working procedure for the endotoxin-free plasmid preparation is identical to the standard protocol and each prep yields up to 500 µg of endotoxin-free plasmid DNA.
NucleoBond EF kits employ a modified alkaline/SDS lysis procedure to prepare the bacterial cell pellet for plasmid purification. The bacterial lysate is cleared by filtration and loaded onto the equilibrated column. No additional incubation step is required for endotoxin removal. After efficient washing of the column, the purified plasmid DNA is eluted from the column.
- Endotoxins are efficiently removed (<0.1 EU/µg DNA)
- No additional incubation is required for endotoxin removal
- Purify 500 µg from 30–100 ml of overnight E. coli culture
- Patented procedure
|Format||Maxi gravity-flow columns|
|Lysate clarification||Folded filters|
|Starting material||30–150 ml|
|Vector size||<300 kb|
|Typical yield||450–520 µg|
|Endotoxin level||<0.1 EU/µg|
|Preparation time||100 min/2 preps|
Purified DNA is suitable for:
- Transfection of highly sensitive cells
- Lentivirus production
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Tip for a successful transfection—remove endotoxins!
Endotoxins are copurified during plasmid preparations from bacterial lysates and affect eukaryotic cell survival, ultimately leading to a lower transfection efficiency. Innovative mini-format technology reduces endotoxins in your plasmid prep, leads to more successful transfections, and simplifies your workflow.View the data Purchase info
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